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1. Two-photon multiplexing bio-imaging using a combination of Eu- and Tb-bioprobes

Author: Virginie Placide, Delphine Pitrat, Alain Duperray, Alexei Grichine, Chantal Andraud, Anh Thy Bui, Olivier Maury, Laboratoire de Chimie - UMR5182 (LC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École normale supérieure - Lyon (ENS Lyon)-Institut de Chimie du CNRS (INC), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Lanthanide, Photons, Luminescent Agents, Microscope, Materials science, Nanotechnology, Multiplexing, Molecular Imaging, law.invention, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, Inorganic Chemistry, Bio imaging, Europium, Two-photon excitation microscopy, law, Cell Line, Tumor, Research studies, Humans, Terbium, ComputingMilieux_MISCELLANEOUS, Excitation
Abstract: During the course of our research studies aiming to combine the unique photophysical properties of lanthanide complexes and the intrinsic advantage of a nonlinear two-photon excitation, we described the proof-of-concept of a biphotonic multiplexing bio-imaging experiment. To that end, a new Tb-luminescent bioprobe [TbL(2)] was designed using the TACN-trispicolinate platform. The biphotonic imaging of T24 fixed cells stained either with [TbL(2)] or with the already described [EuL(1)] complexes is reported. Finally the biphotonic multiplexing experiment was performed using a combination of both probes using the spectral mode of the two-photon microscope.
Published: 2015

2. Mechanosensitivity of Cancer Cells in Contact with Soft Substrates Using AFM

Author: Valentin Laplaud, Claude Verdier, Yara Abidine, Vinoth Sundar Rajan, Alain Duperray, Richard Michel, Valérie M. Laurent, Andrei Constantinescu, LIPHY-DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Laboratoire de mécanique des solides (LMS), École polytechnique (X)-MINES ParisTech - École nationale supérieure des mines de Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), 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 [2016-2019] (UGA [2016-2019]), ANR-12-BS09-0020,TRANSMIG,Migration transendothéliale de cellules cancéreuses(2012), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA), École polytechnique (X)-MINES ParisTech - École nationale supérieure des mines de Paris-Centre National de la Recherche Scientifique (CNRS), Institut Mondor de recherche biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), DYFCOM, Verdier, Claude, BLANC - Migration transendothéliale de cellules cancéreuses - - TRANSMIG2012 - ANR-12-BS09-0020 - BLANC - VALID, DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM-LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), and École polytechnique (X)-Mines Paris - PSL (École nationale supérieure des mines de Paris)
Subjects: 0301 basic medicine, transendothelial migration, Materials science, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Biophysics, Nanotechnology, Microscopy, Atomic Force, Viscoelasticity, 03 medical and health sciences, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Cell Line, Tumor, Microscopy, Humans, Neoplasm Invasiveness, Elasticity (economics), Cytoskeleton, Elastic modulus, viscoelasticity, Mechanical Phenomena, substrate corrections, mechanosenstivity, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Actin cytoskeleton, Elasticity, Biomechanical Phenomena, indentation, 030104 developmental biology, Cell Biophysics, Cancer cell, cancer cells, Mechanosensitive channels, AFM, substrate correction, Rheology
Abstract: International audience; Cancer cells are usually found to be softer than normal cells, but their stiffness changes when they are in contactwith different environments because of mechanosensitivity. For example, they adhere to a given substrate by tuning theircytoskeleton, thus affecting their rheological properties. This mechanism could become efficient when cancer cells invadethe surrounding tissues, and they have to remodel their cytoskeleton in order to achieve particular deformations. Here weuse an atomic force microscope in force modulation mode to study how local rheological properties of cancer cells are affectedby a change of the environment. Cancer cells were plated on functionalized polyacrylamide substrates of different stiffnesses aswell as on an endothelium substrate. A new correction of the Hertz model was developed because measurements require one toaccount for the precise properties of the thin, layered viscoelastic substrates. The main results show the influence of local cellrheology (the nucleus, perinuclear region, and edge locations) and the role of invasiveness. A general mechanosensitive trend isfound by which the cell elastic modulus and transition frequency increase with substrate elasticity, but this tendency breaksdown with a real endothelium substrate. These effects are investigated further during cell transmigration, when the actincytoskeleton undergoes a rapid reorganization process necessary to push through the endothelial gap, in agreement with thelocal viscoelastic changes measured by atomic force microscopy. Taken together, these results introduce a paradigm for anew—to our knowledge—possible extravasation mechanism.
Published: 2017

3. Keto-polymethines: a versatile class of dyes with outstanding spectroscopic properties for in cellulo and in vivo two-photon microscopy imaging

Author: San-Hui Chi, Boudewijn van der Sanden, Olivier Maury, Denis Jacquemin, Chantal Andraud, Joseph W. Perry, Alexei Grichine, Cyrille Monnereau, Alain Duperray, Boris Le Guennic, Jérôme Cuny, Simon Pascal, Florence Appaix, Sandrine Denis-Quanquin, Laboratoire de Chimie - UMR5182 (LC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École normale supérieure - Lyon (ENS Lyon)-Institut de Chimie du CNRS (INC), Grenoble Institut des Neurosciences (GIN), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), LIPHY-DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut des Sciences Chimiques de Rennes (ISCR), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Chimie Et Interdisciplinarité : Synthèse, Analyse, Modélisation (CEISAM), Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Modélisation, Agrégats, Dynamique (LCPQ) (MAD), Laboratoire de Chimie et Physique Quantiques (LCPQ), Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), School of Chemistry and Biochemistry, and Center for Organic Electronics and Photonics, Georgia Institute of Technology [Atlanta], D. J. acknowledges the ERC for financial support in the framework of a Starting Grant (Marches – 278845). J. W. P. and S.-H. C. acknowledge the support from the DARPA ZOE Program (W31P4Q-09-1-0012), and the AFOSR MURI (FA9550-10-1-0558)., Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Nantes - UFR des Sciences et des Techniques (UN UFR ST), Université de Nantes (UN)-Université de Nantes (UN), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM-LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Nationale Supérieure de Chimie de Rennes (ENSCR)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Nantes (UN)-Université de Nantes (UN)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Elefantis, Nicolas, École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), [GIN] Grenoble Institut des Neurosciences (GIN), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])
Subjects: Chemistry, Stereochemistry, Hydrogen bond, Quantum yield, 02 engineering and technology, General Chemistry, 010402 general chemistry, 021001 nanoscience & nanotechnology, Photochemistry, 01 natural sciences, Fluorescence, 0104 chemical sciences, Molecular engineering, [CHIM.THEO]Chemical Sciences/Theoretical and/or physical chemistry, [CHIM.THEO] Chemical Sciences/Theoretical and/or physical chemistry, Molecular dynamics, Membrane, Two-photon excitation microscopy, Molecule, 0210 nano-technology
Abstract: International audience; The synthesis of keto-heptamethine derivatives has been expanded to various new symmetrical and asymmetrical structures, including an unprecedented di-anionic keto-polymethine. The spectroscopic behavior of these new dyes has been systematically and thoroughly investigated, revealing that the formation of hydrogen bond interactions with protic solvents is responsible for a dramatic enhancement of the fluorescence quantum yield in the far-red spectral region. The existence of these strong hydrogen-bond interactions was further confirmed by molecular dynamics simulations. These bis-dipolar polymethines exhibit large two-photon absorption (TPA) cross-sections (σ2 in GM) in the near-infrared, making them ideal candidates for NIR-to-NIR two-photon microscopy imaging applications. We demonstrate that the molecular engineering of the hydrophilic/hydrophobic balance enables targeting of different cellular components, such as cytoplasm or cell membranes. Addition of appropriate substituents provides the molecule with high-water-solubility, affording efficient two-photon probes for angiography.
Published: 2017

4. Mathematical framework for traction force microscopy

Author: Guido Vitale, Claude Verdier, Valentina Peschetola, Jocelyn Etienne, Alain Duperray, Luigi Preziosi, Richard Michel, Davide Carlo Ambrosi, DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire de mécanique des solides (LMS), École polytechnique (X)-MINES ParisTech - École nationale supérieure des mines de Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), MOX, Milano, MOX-Dipartimento di Matematica (MOX), Politecnico di Milano [Milan] (POLIMI)-Politecnico di Milano [Milan] (POLIMI), Matematica, Politecnico di Torino, Dipartimento di Matematica 'Giuseppe Peano' [Torino], and Università degli studi di Torino (UNITO)-Università degli studi di Torino (UNITO)
Subjects: Surface (mathematics), Inverse problems, Mathematical optimization, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Traction (engineering), Living cell, Cell motility, 01 natural sciences, Traction force microscopy, Quantitative Biology::Cell Behavior, Tikhonov regularization, 03 medical and health sciences, symbols.namesake, Adjoint Method (AM), [MATH.MATH-MP]Mathematics [math]/Mathematical Physics [math-ph], QA1-939, 0101 mathematics, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], 030304 developmental biology, Mathematics, 0303 health sciences, T57-57.97, Applied mathematics. Quantitative methods, Mathematical analysis, Minimization problem, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Inverse problem, 010101 applied mathematics, Fourier transform, symbols, L -curve, Fourier Transform Traction Cytometry (FTTC)
Abstract: This paper deals with the Traction Force Microscopy (TFM) problem. It consists in obtaining stresses by solving an inverse problem in an elastic medium, from known experimentally measured displacements. In this article, the application is the determination of the stresses exerted by a living cell at the surface of an elastic gel. We propose an abstract framework which formulates this inverse problem as a constrained minimization one. The mathematical constraints express the biomechanical conditions that the stress field must satisfy. From this framework, two methods currently used can be derived, the adjoint method (AM) and the Fourier Transform Traction Cytometry (FTTC) method. An improvement of the FTTC method is also derived using this framework. The numerical results are compared and show the advantage of the AM, in particular its ability to capture details more accurately. Cet article est consacré au problème de la Microscopie à Force de Traction (TFM). Ce problème consiste à déterminer les contraintes exercées par une cellule lors de sa migration sur un substrat élastique à partir d’une mesure expérimentale des déplacements induits dans ce substrat. Mathématiquement, il s’agit de résoudre un problème inverse pour lequel nous proposons une formulation abstraite de type optimisation sous contraintes. Les contraintes mathématiques expriment les constraintes biomécaniques que doit satisfaire le champ de contraintes exercé par la cellule. Ce cadre abstrait permet de retrouver deux des méthodes de résolution utilisées en pratique, à savoir la méthode adjointe (AM) et la méthode de Cytométrie de Traction par Transformée de Fourier (FTTC). Il permet aussi d’ameliorer la méthode FTTC. Les résultats numériques obtenus sont ensuite comparés et démontrent l’avantage de la méthode adjointe, en particulier par sa capacité à capturer des détails avec une meilleure précision.
Published: 2013

5. The impact of cardiac ischemia/reperfusion on the mitochondria-cytoskeleton interactions

Author: Stéphane Tanguy, Karin Pernet-Gallay, Rafaela Bagur, Sarah Foriel, Alexei Grichine, Tuuli Kaambre, François Boucher, Rita Guzun, Caroline Sanchez, Yves Usson, Andrey V. Kuznetsov, Laboratory of Fundamental and Applied Bioenergetics = Laboratoire de bioénergétique fondamentale et appliquée (LBFA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Physiologie cardio-Respiratoire Expérimentale Théorique et Appliquée (TIMC-IMAG-PRETA), 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]), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), LIPHY-DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Grenoble Institut des Neurosciences (GIN), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Cardiac Surgery Research Laboratory, Department of Heart Surgery-Innsbruck Medical University [Austria] (IMU), Dynamique Cellulaire et Tissulaire- Interdisciplinarité, Modèles & Microscopies (TIMC-IMAG-DyCTiM), and Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: 0301 basic medicine, Male, [SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging, Heart Ventricles, Cell Respiration, Myocardial Reperfusion Injury, Oxidative phosphorylation, Mitochondrion, Creatine, Mitochondria, Heart, Phosphocreatine, 03 medical and health sciences, chemistry.chemical_compound, Tubulin, Animals, Rats, Wistar, Cytoskeleton, Molecular Biology, 030102 biochemistry & molecular biology, biology, Myocardium, Cell biology, 030104 developmental biology, chemistry, biology.protein, Molecular Medicine, Creatine kinase, Intracellular
Abstract: International audience; Cardiac ischemia-reperfusion (IR) injury compromises mitochondrial oxidative phosphorylation (OxPhos) and compartmentalized intracellular energy transfer via the phosphocreatine/creatine kinase (CK) network. The restriction of ATP/ADP diffusion at the level of the mitochondrial outer membrane (MOM) is an essential element of compartmentalized energy transfer. In adult cardiomyocytes, the MOM permeability to ADP is regulated by the interaction of voltage-dependent anion channel with cytoskeletal proteins, particularly with β tubulin II. The IR-injury alters the expression and the intracellular arrangement of cytoskeletal proteins. The objective of the present study was to investigate the impact of IR on the intracellular arrangement of β tubulin II and its effect on the regulation of mitochondrial respiration. Perfused rat hearts were subjected to total ischemia (for 20min (I20) and 45min (I45)) or to ischemia followed by 30min of reperfusion (I20R and I45R groups). High resolution respirometry and fluorescent confocal microscopy were used to study respiration, β tubulin II and mitochondrial arrangements in cardiac fibers. The results of these experiments evidence a heterogeneous response of mitochondria to IR-induced damage. Moreover, the intracellular rearrangement of β tubulin II, which in the control group colocalized with mitochondria, was associated with increased apparent affinity of OxPhos for ADP, decreased regulation of respiration by creatine without altering mitochondrial CK activity and the ratio between octameric to dimeric isoenzymes. The results of this study allow us to highlight changes of mitochondrial interactions with cytoskeleton as one of the possible mechanisms underlying cardiac IR injury.
Published: 2016

6. Blood flow and microgravity

Author: Chaouqi Misbah, Lionel Bureau, Alain Duperray, Thomas Podgorski, Daria Tsvirkun, Mikhail Vysokikh, Alexander Farutin, Frank Dubois, Gwennou Coupier, Christophe Minetti, DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM-LIPhy), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), LITT&ARTS. Arts et pratiques du texte, de l’image, de l’écran et de la scène (LITT&ARTS ), LIPHY-DYFCOM, Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Physique et mécanique des milieux hétérogenes (UMR 7636) (PMMH), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), and Research Center for Obstetrics, Gynecology and Perinatology [Moscow, Russian Federation]
Subjects: Polymer brush, Endothelium, Strategy and Management, Physical activity, Hemodynamics, Mechanical engineering, 01 natural sciences, 010305 fluids & plasmas, Glycocalyx, Materials Science(all), Écoulement sanguin, 0103 physical sciences, Media Technology, medicine, Brosse de polymère, General Materials Science, Endothélium, Mechanotransduction, 010306 general physics, Force de portance, Marketing, Chemistry, Collective motion, Blood flow, Microgravité, Lift force, medicine.anatomical_structure, Mechanics of Materials, Biophysics, Microgravity, [PHYS.COND.CM-SCM]Physics [physics]/Condensed Matter [cond-mat]/Soft Condensed Matter [cond-mat.soft]
Abstract: International audience; The absence of gravity during space flight can alter cardio-vascular functions partially due to reduced physical activity. This affects the overall hemodynamics, and in particular the level of shear stresses to which blood vessels are submitted. Long-term exposure to space environment is thus susceptible to induce vascular remodeling through a mechanotransduction cascade that couples vessel shape and function with the mechanical cues exerted by the circulating cells on the vessel walls. Central to such processes, the glycocalyx – i.e. the micron-thick layer of biomacromolecules that lines the lumen of blood vessels and is directly exposed to blood flow – is a major actor in the regulation of biochemical and mechanical interactions. We discuss in this article several experiments performed under microgravity, such as the determination of lift force and collective motion in blood flow, and some preliminary results obtained in artificial microfluidic circuits functionalized with endothelium that offer interesting perspectives for the study of the interactions between blood and endothelium in healthy condition as well as by mimicking the degradation of glycocalyx caused by long space missions. A direct comparison between experiments and simulations is discussed.
Published: 2016

7. Chauffage par effet plasmon pour la désorption localisée de cellules capturées sur une biopuce

Author: Thierry Livache, Yoann Roupioz, Dieudonné R. Baganizi, Patrice N. Marche, Roberto Calemczuk, Radoslaw Bombera, Elodie Engel, Loïc Leroy, Loïc Laplatine, Structures et propriétés d'architectures moléculaire (SPRAM - UMR 5819), Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut d'oncologie/développement Albert Bonniot de Grenoble, Université Joseph Fourier - Grenoble 1 (UJF)-CHU GRENOBLE-Inserm : U823-EFS, Chimie pour la Reconnaissance et l’Etude d’Assemblages Biologiques (CREAB), SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé (SYMMES), Institut de Chimie du CNRS (INC)-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)-Institut de Chimie du CNRS (INC)-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), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Chimie du CNRS (INC)-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)-Institut de Chimie du CNRS (INC)-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), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), CHU GRENOBLE-Université Joseph Fourier - Grenoble 1 (UJF)-Inserm : U823-EFS, Engel, Elodie, and Sciencesconf.org, CCSD
Subjects: [PHYS]Physics [physics], [SPI] Engineering Sciences [physics], Chemistry, Analytical chemistry, 02 engineering and technology, 010402 general chemistry, 021001 nanoscience & nanotechnology, 01 natural sciences, [PHYS] Physics [physics], 0104 chemical sciences, [SPI]Engineering Sciences [physics], Biophysics, Laser illumination, Surface plasmon resonance, 0210 nano-technology, Biochip, Engineering (miscellaneous), Instrumentation, ComputingMilieux_MISCELLANEOUS
Abstract: International audience; This work proposes a miniaturized system able to perform multiple cell capture followed by cell-type selective release from a biochip surface. Unlabeled lymphocytes are first specifically captured onto a DNA array by antibody-DNA conjugates. The immobilized cells are subsequently released under spatio-temporal control within local heating generated by intense Surface Plasmon Resonance (SPR) produced by laser illumination.
Published: 2016

8. Prediction of traction forces of motile cells

Author: Alain Duperray, Richard Michel, Claude Verdier, Jocelyn Etienne, Clément Roux, Valentina Peschetola, Valérie M. Laurent, Etienne, Jocelyn, BLANC - Migration transendothéliale de cellules cancéreuses - - TRANSMIG2012 - ANR-12-BS09-0020 - BLANC - VALID, DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM-LIPhy), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), ANR-12-BS09-0020,TRANSMIG,Migration transendothéliale de cellules cancéreuses(2012), LIPHY-DYFCOM, Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: 0301 basic medicine, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Traction (engineering), Biomedical Engineering, Biophysics, Bioengineering, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Biochemistry, Single frame, Traction force microscopy, Biomaterials, [PHYS.MECA.MEMA]Physics [physics]/Mechanics [physics]/Mechanics of materials [physics.class-ph], 03 medical and health sciences, Rheology, [PHYS.MECA.MEMA] Physics [physics]/Mechanics [physics]/Mechanics of materials [physics.class-ph], [SDV.BC.BC] Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], [PHYS.MECA.MEFL] Physics [physics]/Mechanics [physics]/Fluid mechanics [physics.class-ph], [PHYS.MECA.MEFL]Physics [physics]/Mechanics [physics]/Fluid mechanics [physics.class-ph], Simulation, Physics, Tractive force, [PHYS.PHYS.PHYS-BIO-PH] Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Mechanics, Articles, Finite element method, 030104 developmental biology, Adhesive, Biotechnology
Abstract: When crawling on a flat substrate, living cells exert forces on it via adhesive contacts, enabling them to build up tension within their cytoskeleton and to change shape. The measurement of these forces has been made possible by traction force microscopy (TFM), a technique which has allowed us to obtain time-resolved traction force maps during cell migration. This cell ‘footprint’ is, however, not sufficient to understand the details of the mechanics of migration, that is how cytoskeletal elements (respectively, adhesion complexes) are put under tension and reinforce or deform (respectively, mature and/or unbind) as a result. In a recent paper, we have validated a rheological model of actomyosin linking tension, deformation and myosin activity. Here, we complement this model with tentative models of the mechanics of adhesion and explore how closely these models can predict the traction forces that we recover from experimental measurements during cell migration. The resulting mathematical problem is a PDE set on the experimentally observed domain, which we solve using a finite-element approach. The four parameters of the model can then be adjusted by comparison with experimental results on a single frame of an experiment, and then used to test the predictive power of the model for following frames and other experiments. It is found that the basic pattern of traction forces is robustly predicted by the model and fixed parameters as a function of current geometry only.
Published: 2016

9. Wavelet transform analysis of chromatin texture changes during heat shock

Author: Herbomel, A., Grichine, A., Arnold Fertin, Delon, A., C. Vourc'H, Souchier, C., Usson, Y., INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), LIPHY-DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Dynamique Cellulaire et Tissulaire- Interdisciplinarité, Modèles & Microscopies (TIMC-IMAG-DyCTiM), 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), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), ANR-08-PCVI-0004,Dynastress,Etude, par microscopie à corrélation de fluorescence, de la réponse nucléaire au stress thermique, and aspects dynamiques.(2008)
Subjects: Microscopy, Confocal, Heat Shock Transcription Factors, Cell Survival, [SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging, Image Processing, Computer-Assisted, Wavelet Analysis, Humans, Algorithms, Chromatin, Heat-Shock Response, ComputingMilieux_MISCELLANEOUS, HeLa Cells
Abstract: Texture analysis can be a useful tool to investigate the organization of chromatin. Approaches based on multiscale analysis and in particular the 'à trou' wavelet analysis has already been used for microscopy (Olivo Marin). In order to analyse texture changes, the statistical properties of the wavelet coefficient images were summarized by the first four statistical orders: mean, standard deviation, skewness and kurtosis of the coefficient image histogram. The 'à trou' transform provided a representation of the wavelet coefficients and texture parameters with the same statistical robustness throughout the scale spaces. It was applied for quantifying chromatin texture and heat-induced chromatin changes in living cells. We investigated the changes by both laser scanning and spinning disk confocal microscopies and compared the texture parameters before and after increasing duration of heat shock exposure (15 min, 30 min and 1 h). Furthermore, as activation of the heat shock response also correlates with a rapid localization of HSF1 within a few nuclear structures termed nuclear stress bodies (nSBs), we compared the dynamics of nSBs formation with that of textural changes during 1 h of continuous heat shock. Next, we studied the recovery phase following a 1-h heat shock. Significant differences were observed, particularly affecting the perinucleolar region, even for the shortest heat shock time affecting mostly the skewness and standard deviation. Furthermore, progressive changes could be observed according to the duration of heat shock, mostly affecting fine details (pixel-wise changes) as revealed by the parameters, obtained from the first- and second-order wavelet coefficients. 'A trou' wavelet texture analysis provided a sensitive and efficient tool to investigate minute changes of chromatin.
Published: 2015

10. 3D cancer cell migration in collagen matrices

Author: Laure Laforgue, Alain Duperray, Claude Verdier, Valérie M. Laurent, DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), SB, and ANR-12-BS09-0020,TRANSMIG,Migration transendothéliale de cellules cancéreuses(2012)
Subjects: collagen, velocity, [SDV]Life Sciences [q-bio], Cell, Biomedical Engineering, Motility, Bioengineering, Biology, Matrix (biology), migration, 03 medical and health sciences, MSD, 0302 clinical medicine, medicine, cancer, Cytoskeleton, 030304 developmental biology, [PHYS]Physics [physics], 0303 health sciences, Cancer, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Cell migration, General Medicine, Adhesion, medicine.disease, Computer Science Applications, Human-Computer Interaction, medicine.anatomical_structure, Cancer cell, Biophysics, 030217 neurology & neurosurgery, Biomedical engineering
Abstract: International audience; Cell migration is a widely studied subject but often limited to 2D motion (Sheetz et al., 1999) on various substrates for it allows to simplify cell-substrate interactions. The important relevant parameters to 2D cell migration are substrate rigidity, adhesion, and the role of the cell cytoskeleton. More recently, new studies on 3D cell migration have been performed thanks to the development of new visualization tools such as reflectance confocal microscopy (Iordan et al. 2010, Wolf et al. 2013). In particular, it is possible to image cell and the extra-cellular matrix at the same time. This leads to interesting applications on cancer cell migration, and could possibly open new routes to the understanding of cancer development, like the mechanisms used by cancer cells to invade new tissues.Thus, the aim of this study is to study cancer migration in collagen matrices, as a model of soft tissue, to investigate their motility within the surrounding collagen matrix. Different collagen concentrations will be used and the ability of these cancer cells to move through such a complex structure will be addressed. To this aim, first results of cell migration velocity as well as the Mean Squared Displacement (MSD) will be analyzed.
Published: 2015

11. Innovative immuno-assay exploiting magnetic nanoparticles and structured micro-magnets

Author: Delshadi, Sarah, Blaire, Guillaume, Kauffmann, Paul, Fratzl, Mario, Devillers, Thibaut, Reyne, Gilbert, Delabouglise, Didier, Weindenhaupt, Marianne, Dempsey, Nora M., Bruckert, Franz, Marche, Patrice, Cugat, Orphée, Laboratoire de Génie Electrique de Grenoble (G2ELab), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut Polytechnique de Grenoble - Grenoble Institute of Technology-Centre National de la Recherche Scientifique (CNRS), Laboratoire Biopuces (BIOPUCES), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Micro et NanoMagnétisme (NEEL - MNM), Institut Néel (NEEL), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS), Laboratoire des matériaux et du génie physique (LMGP ), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut National Polytechnique de Grenoble (INPG)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Micro et NanoMagnétisme (MNM), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Institut National Polytechnique de Grenoble (INPG)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: [SPI.NRJ]Engineering Sciences [physics]/Electric power
Abstract: International audience; Magnetic attraction is widely used in biotechnology, because it provides long-range forces able to capture molecules of interest.Micro-magnet arrays from Néel Institute (CNRS, Grenoble, FRANCE) can efficiently trap highly diffusive magnetic nanoparticles thanksto their high local gradients. This innovative technology is used to develop fast immuno-assays that take advantage of a radical sizereduction, compared to commercial technology.Our first objective is to embed micro-magnets in 96 wells plates to accelerate and enhance performances of standard ELISA.As micro-magnets are embedded in standard microwell plates, our plates remain fully compatible with plates readers. We willcompare the ELISA performances applied to ovalbumin antibody detection.
Published: 2015

12. Unexpected Efficiency of a Luminescent Samarium(III) Complex for Combined Visible and Near-Infrared Biphotonic Microscopy

Author: Anh Thy Bui, Olivier Maury, Alain Duperray, Alexei Grichine, Sophie Brasselet, Chantal Andraud, Laboratoire de Chimie - UMR5182 (LC), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut FRESNEL (FRESNEL), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS), MOSAIC (MOSAIC), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Groupe de Recherche Sur Le Cancer du Poumon : Bases Moléculaires de la Progression Tumorale, Dépistage et Thérapie Génique, Institut Albert Bonniot-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Albert Bonniot-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École normale supérieure - Lyon (ENS Lyon)-Institut de Chimie du CNRS (INC), Centre National de la Recherche Scientifique (CNRS)-École Centrale de Marseille (ECM)-Aix Marseille Université (AMU), and Centre National de la Recherche Scientifique (CNRS)-École Centrale de Marseille (ECM)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Marseille (ECM)-Aix Marseille Université (AMU)
Subjects: Lanthanide, Brightness, Organic Chemistry, Near-infrared spectroscopy, Analytical chemistry, chemistry.chemical_element, Astrophysics::Cosmology and Extragalactic Astrophysics, General Chemistry, Chromophore, Photochemistry, Catalysis, Samarium, chemistry, Microscopy, [SPI.OPTI]Engineering Sciences [physics]/Optics / Photonic, Luminescence, Astrophysics::Galaxy Astrophysics, ComputingMilieux_MISCELLANEOUS
Abstract: An original samarium(III) complex based on a triazacyclononane platform functionalized with a charge-transfer antenna chromophore exhibited optimized brightness and was successfully used as an emissive species for two-photon microscopy experiments in both the visible and near-infrared spectral ranges.
Published: 2015

13. CCM proteins control endothelial b1 integrin dependent response to shear stress

Author: Claude Verdier, Valérie Deplano, Eva Faurobert, Justyna Lisowska, Corinne Albiges-Rizo, Alain Duperray, Christian Geindreau, Zuzana Macek Jilkova, Sandra Manet, INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Dynamique des systèmes d'adhérence et différenciation (DySAD), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherche sur les Phénomènes Hors Equilibre (IRPHE), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire sols, solides, structures - risques [Grenoble] (3SR), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS), Mécanique et Couplages Multiphysiques des Milieux Hétérogènes (CoMHet), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS), LIPHY-DYFCOM, Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-École Centrale de Marseille (ECM), Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS), and Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Endothelium, Mechanotransduction, QH301-705.5, Science, [SDV]Life Sciences [q-bio], Cell, Biology, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, 0302 clinical medicine, β1 integrin, medicine, Shear stress, Extracellular, Biology (General), Cytoskeleton, Actin, 030304 developmental biology, CCM, [PHYS]Physics [physics], 0303 health sciences, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Cell biology, medicine.anatomical_structure, b1 integrin, Immunology, General Agricultural and Biological Sciences, 030217 neurology & neurosurgery, Intracellular, Research Article
Abstract: Hemodynamic shear stress from blood flow on the endothelium critically regulates vascular function in many physiological and pathological situations. Endothelial cells adapt to shear stress by remodeling their cytoskeletal components and subsequently by changing their shape and orientation. We demonstrate that β1 integrin activation is critically controlled during the mechanoresponse of endothelial cells to shear stress. Indeed, we show that overexpression of the CCM complex, an inhibitor of β1 integrin activation, blocks endothelial actin rearrangement and cell reorientation in response to shear stress similarly to β1 integrin silencing. Conversely, depletion of CCM2 protein leads to an elongated “shear-stress-like” phenotype even in the absence of flow. Taken together, our findings reveal the existence of a balance between positive extracellular and negative intracellular signals, i.e. shear stress and CCM complex, for the control of β1 integrin activation and subsequent adaptation of vascular endothelial cells to mechanostimulation by fluid shear stress.
Published: 2014

14. Design and synthesis of europium luminescent bio-probes featuring sulfobetaine moieties

Author: Virginie Placide, Delphine Pitrat, Alexei Grichine, Alain Duperray, Chantal Andraud, Olivier Maury, Laboratoire de Chimie - UMR5182 (LC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École normale supérieure - Lyon (ENS Lyon)-Institut de Chimie du CNRS (INC), LIPHY-DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), and Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Lanthanide, Aqueous solution, Ligand, Chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, chemistry.chemical_element, [CHIM.CATA]Chemical Sciences/Catalysis, [CHIM.MATE]Chemical Sciences/Material chemistry, [CHIM.INOR]Chemical Sciences/Inorganic chemistry, Biochemistry, Combinatorial chemistry, Non specific, Drug Discovery, Organic chemistry, Macrocyclic ligand, Luminescence, Europium, ComputingMilieux_MISCELLANEOUS, Europium luminescence
Abstract: Herein we report the straightforward preparation of chromophore-functionalized TACN ligand via Cu-free cross-coupling reactions using a common halogenated platform. This versatile methodology allows the preparation of original macrocyclic ligand featuring both optimized antenna for the sensitization of europium luminescence and sulfobetaine zwitterionic groups to ensure water solubility of the complex. In addition preliminary two-photon excited microscopy imaging experiments of fixed cells reveal that sulfobetaine groups are able to limit undesirable non specific interactions with biological surrounding.
Published: 2014

15. Atomic Force microscopy reveals a role for endothelial cell ICAM-1 expression in bladder cancer cell adherence

Author: Alain Duperray, Claude Verdier, Vinoth Sundar Rajan, Valérie M. Laurent, Verdier, Claude, DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), DYFCOM, Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), and Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: Ligands, Microscopy, Atomic Force, Biochemistry, Physical Chemistry, Forces, Epithelium, 0302 clinical medicine, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Molecular Cell Biology, Basic Cancer Research, Medicine and Health Sciences, CD43, Biomacromolecule-Ligand Interactions, Cancer, 0303 health sciences, ICAM-1, Multidisciplinary, Leukosialin, biology, [PHYS.PHYS.PHYS-BIO-PH] Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Chemistry, Physics, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Intercellular Adhesion Molecule-1, Bladder Cancer, Invasiveness, Cell biology, Biomechanical Phenomena, Endothelial stem cell, Gene Expression Regulation, Neoplastic, medicine.anatomical_structure, Oncology, 030220 oncology & carcinogenesis, Physical Sciences, Adhesion, Medicine, Anatomy, Cellular Types, AFM, Research Article, Endothelium, Science, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Integrin, Biophysics, Receptor-ligand, MUC1, 03 medical and health sciences, Cell Line, Tumor, medicine, Cell Adhesion, Humans, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Cell adhesion, 030304 developmental biology, Intermolecular Forces, Mucin-1, [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Biology and Life Sciences, Endothelial Cells, Cancers and Neoplasms, Epithelial Cells, Cell Biology, medicine.disease, Genitourinary Tract Tumors, Biological Tissue, Urinary Bladder Neoplasms, Chemical Properties, Cell culture, Cancer cell, biology.protein
Abstract: International audience; Cancer metastasis is a complex process involving cell-cell interactions mediated by cell adhesive molecules. In this study we determine the adhesion strength between an endothelial cell monolayer and tumor cells of different metastatic potentials using Atomic Force Microscopy. We show that the rupture forces of receptor-ligand bonds increase with retraction speed and range between 20 and 70 pN. It is shown that the most invasive cell lines (T24, J82) form the strongest bonds with endothelial cells. Using ICAM-1 coated substrates and a monoclonal antibody specific for ICAM-1, we demonstrate that ICAM-1 serves as a key receptor on endothelial cells and that its interactions with ligands expressed by tumor cells are correlated with the rupture forces obtained with the most invasive cancer cells (T24, J82). For the less invasive cancer cells (RT112), endothelial ICAM-1 does not seem to play any role in the adhesion process. Moreover, a detailed analysis of the distribution of rupture forces suggests that ICAM-1 interacts preferentially with one ligand on T24 cancer cells and with two ligands on J82 cancer cells. Possible counter receptors for these interactions are CD43 and MUC1, two known ligands for ICAM-1 which are expressed by these cancer cells.
Published: 2014

16. 3D migration of cells solving an inverse problem

Author: Vitale, Guido, Laforgue, Laure, Michel, Richard, Etienne, Jocelyn, Laurent, Valérie M., Duperray, Alain, Verdier, Claude, Verdier, Claude, BLANC - Migration transendothéliale de cellules cancéreuses - - TRANSMIG2012 - ANR-12-BS09-0020 - BLANC - VALID, DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Groupe de Recherche Sur Le Cancer du Poumon : Bases Moléculaires de la Progression Tumorale, Dépistage et Thérapie Génique, Institut Albert Bonniot-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Albert Bonniot-Institut National de la Santé et de la Recherche Médicale (INSERM), and ANR-12-BS09-0020,TRANSMIG,Migration transendothéliale de cellules cancéreuses(2012)
Subjects: [PHYS]Physics [physics], [SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Quantitative Biology::Cell Behavior, [PHYS] Physics [physics]
Abstract: International audience; Traction Force Microscopy (TFM) is an inverse method that allows to obtain the stress field applied by a living cell on the environment on the basis of a pointwise knowledge of the displacement produced by the cell itself during its migration. This biophysical problem, usually addressed in terms of Green functions, can be alternatively tackled in a variational framework. In such a case, a suitable penalty functional has to be minimized. The resulting Euler-Lagrange equations include both the direct problem based on the linear elasticity operator as well as an equation built on its adjoint. Results from a two-dimensional model, i.e. where living cancer cells are migrating on a plane substrate, are briefly presented. While the mathematics is well established also in the three-dimensional case, i.e. where cells are completely embedded in the gel matrix, the experimental data needed are more difficult to obtain than the two-dimensional counterpart. First steps towards the complete three-dimensional traction reconstruction are reported.
Published: 2014

17. Amoeboid Swimming: A Generic Self-Propulsion of Cells in Fluids by Means of Membrane Deformations

Author: Philippe Peyla, Alexander Farutin, Chaouqi Misbah, Dag Kristian Dysthe, Alain Duperray, Salima Rafaï, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Physics of Geological Processes [Oslo] (PGP), Department of Physics [Oslo], Faculty of Mathematics and Natural Sciences [Oslo], University of Oslo (UiO)-University of Oslo (UiO)-Faculty of Mathematics and Natural Sciences [Oslo], University of Oslo (UiO)-University of Oslo (UiO)-Department of Geosciences [Oslo], University of Oslo (UiO)-University of Oslo (UiO), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Farutin, Alexander, Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), and Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Physics, Linear function (calculus), Euglenida, Force density, [PHYS.PHYS.PHYS-BIO-PH] Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], General Physics and Astronomy, Nanotechnology, Mechanics, Propulsion, Flagellum, 01 natural sciences, Models, Biological, 010305 fluids & plasmas, Membrane, 47.63.Gd, 47.15.G-, 47.61.-k, 0103 physical sciences, Compressibility, Torque, Cell Surface Extensions, 010306 general physics, active swimming, Swimming, Dimensionless quantity, microswimmers
Abstract: International audience; Microorganisms, such as bacteria, algae, or spermatozoa, are able to propel themselves forward thanks to flagella or cilia activity. By contrast, other organisms employ pronounced changes of the membrane shape to achieve propulsion, a prototypical example being the Eutreptiella gymnastica. Cells of the immune system as well as dictyostelium amoebae, traditionally believed to crawl on a substratum, can also swim in a similar way. We develop a model for these organisms: the swimmer is mimicked by a closed incompressible membrane with force density distribution (with zero total force and torque). It is shown that fast propulsion can be achieved with adequate shape adaptations. This swimming is found to consist of an entangled pusher-puller state. The auto-propulsion distance over one cycle is a universal linear function of a simple geometrical dimensionless quantity $A/V^{2/3}$ (V and A are cell volume and its membrane area). This study captures the peculiar motion of Eutreptiella gymnastica with simple force distribution.
Published: 2013

18. Wall shear stress and endothelial cells dysfunction in the context of abdominal aortic aneurysms

Author: Valérie Deplano, Z. Macek Jilkova, Claude Verdier, Alain Duperray, Christian Geindreau, Mamadou Toungara, Verdier, Claude, 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), Institut de Recherche sur les Phénomènes Hors Equilibre (IRPHE), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Laboratoire sols, solides, structures - risques [Grenoble] (3SR), Université Joseph Fourier - Grenoble 1 (UJF)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-École Centrale de Marseille (ECM), DYFCOM, and Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP )-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National Polytechnique de Grenoble (INPG)-Centre National de la Recherche Scientifique (CNRS)
Subjects: medicine.medical_specialty, Umbilical Veins, Biomedical Engineering, Hemodynamics, Bioengineering, Context (language use), 030204 cardiovascular system & hematology, Wall shear, 03 medical and health sciences, 0302 clinical medicine, Aneurysm, medicine.artery, Internal medicine, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], medicine, Shear stress, Humans, Aorta, Abdominal, Mechanotransduction, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], 030304 developmental biology, 0303 health sciences, Analysis of Variance, business.industry, Abdominal aorta, [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Models, Cardiovascular, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Endothelial Cells, General Medicine, Anatomy, medicine.disease, Abdominal aortic aneurysm, Computer Science Applications, Human-Computer Interaction, Cardiology, Stress, Mechanical, business, Aortic Aneurysm, Abdominal
Abstract: International audience; The formation of aneurysm of abdominal aorta (AAA) is a multifactorial and predominantly degenerative process that results from a complex interplay between biological processes in the arterial wall and the haemodynamic stimuli on the wall, i.e. wall shear stresses (WSSs) and elongations of the wall directly applied on vascular endothelial cells (ECs) partly regulate the arterial wall remodelling (Golledge et al. 2006; Humphrey and Holzapfel 2012). In the case of AAA, these mechanical stimuli strongly vary in space and time, leading to strong spatiotemporal gradients (Figure 1). The effect of these abnormal mechanical stimuli leads to EC dysfunction. This phenomenon plays a significant role in AAA pathology, in which damage of ECs, associated with changes in endothelial permeability and ...
Published: 2013

19. Microrheology of complex systems and living cells using AFM

Author: Yara Abidine, Alain Duperray, Richard Michel, Valérie M. Laurent, Claude Verdier, DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), and Verdier, Claude
Subjects: Microrheology, Materials science, Cantilever, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Microfluidics, Biomedical Engineering, Atomic force acoustic microscopy, Modulus, Bioengineering, Viscoelastic Substances, 02 engineering and technology, Microscopy, Atomic Force, Models, Biological, Viscoelasticity, Cell Physiological Phenomena, Quantitative Biology::Cell Behavior, Shear modulus, 03 medical and health sciences, Optics, Rheology, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Indentation, Viscoelastic properties, Humans, Force modulation mode, Composite material, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], 030304 developmental biology, 0303 health sciences, [PHYS.PHYS.PHYS-BIO-PH] Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], business.industry, [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], technology, industry, and agriculture, polyacrylamide gel, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], General Medicine, 021001 nanoscience & nanotechnology, Computer Science Applications, Atomic Force Microscopy, Human-Computer Interaction, cells, 0210 nano-technology, business
Abstract: International audience; Biological cells display viscoelastic mechanical properties that are a key factor in the regulation of cell processes, such as migration, adhesion and deformability. One interesting tool to probe the mechanical properties of living cells and other complex systems is the Atomic Force Microscopy (AFM). Rheological properties of the sample are obtained usually from the force-indentation (F-δ) measurements and by fitting with the Sneddon's modification of the Hertz model. This model describes the relationship between the force applied by a stiff cone, a purely elastic indentation in a flat and soft sample and the Young's modulus E. AFM uses a flexible cantilever with a pyramidal tip (or sphere) at the end that will locally indent the viscoelastic material. Another approach is to superpose low-amplitude sinusoidal oscillations to an initial identation δ0 (polyacrylamide gel or cells). Then one can determine the complex shear modulus G*...
Published: 2013

20. Time-dependent traction force microscopy for cancer cells as a measure of invasiveness

Author: Peschetola, Valentina, Laurent, Valérie, Duperray, Alain, Michel, Richard, Ambrosi, Davide, Preziosi, Luigi, Verdier, Claude, DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), DYFCOM, INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), MOX, MOX-Dipartimento di Matematica (MOX), Politecnico di Milano [Milan] (POLIMI)-Politecnico di Milano [Milan] (POLIMI), Matematica, Politecnico di Torino, Dipartimento di Matematica 'Giuseppe Peano' [Torino], Università degli studi di Torino (UNITO)-Università degli studi di Torino (UNITO), and co-tutelle franco-italienne
Subjects: adhesion, MSD, invasiveness, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], traction forces, cancer cells, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], migration
Abstract: International audience; The migration of tumor cells of different invasivity is studied, and traction forces are determined in time on soft substrates (Young modulus 10 kPa). It is found that the outliers of the traction stresses are quite relevant to differentiate different cancer cell lines which are more or less invasive. Here we tested two different epithelial bladder cancer cell lines, one invasive (T24), and a less invasive one (RT112). Invasive cancer cells move in a nearly periodic motion, with peaks in velocity corresponding to higher traction forces exerted on the substrate, whereas the less invasive cell develops almost constant time-dependent traction stresses. The dynamics of focal adhesions as well as cytoskeleton features reveal different mechanisms activated to migrate, depending on their invasiveness. T24 invasive cells show an interconnected cytoskeleton linked to mature adhesion sites, leading to small traction stresses, whereas less invasive cells (RT112) show a less-structured cytoskeleton, unmature adhesions corresponding to higher traction stresses. Migration velocities are smaller in the case of less invasive cells. The MSD (Mean Squared Displacement) shows super-diffusive motions with a higher exponent for the more invasive cancer cells. Further correlations between traction forces and the actin cytoskeleton reveal an unexpected pattern with a large actin rim at the RT112 cell edge where higher forces are colocalized, whereas a more usual cytoskeleton with stress fibers and focal adhesions is found for T24 cancer cells. Thus the method can be an interesting one to differentiate cancer cell invasiveness.
Published: 2013

21. Polymer adhesive surface as flexible generic platform for multiplexed assays biochip production

Author: Patrice Faure, Benjamin P. Corgier, Céline A. Mandon, Patrice N. Marche, Christophe A. Marquette, Gaelle C. Le Goff, Loïc J. Blum, Ophélie I. Berthuy, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), AXO Science (Axo S), Hypoxie et physiopathologies cardiovasculaire et respiratoire, Institut National de la Santé et de la Recherche Médicale (INSERM), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Université de Lyon-Université de Lyon-École Supérieure de Chimie Physique Électronique de Lyon (CPE)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Hypoxie : Physiopathologie Respiratoire et Cardiovasculaire (HP2), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Marche, Patrice, Centre National de la Recherche Scientifique (CNRS)-École Supérieure Chimie Physique Électronique de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, HP2, Pole Biol CHU Grenoble (INSERM, U1042), and Université Grenoble Alpes [2016-2019] (UGA [2016-2019])
Subjects: Materials science, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV.IMM] Life Sciences [q-bio]/Immunology, Polymers, [SDV]Life Sciences [q-bio], AFP, Biomedical Engineering, Biophysics, Oligonucleotides, Nanotechnology, 02 engineering and technology, Microarray, 01 natural sciences, Multiplexing, Sensitivity and Specificity, Adhesives, Electrochemistry, Humans, Multiplexed assays, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, Biochip, Localised cell culture, Adhesive surface, Immunoassay, 010401 analytical chemistry, Liver Neoplasms, General Medicine, Adhesion, Equipment Design, 021001 nanoscience & nanotechnology, Microarray Analysis, Hepatitis C, 3. Good health, 0104 chemical sciences, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, [INFO.INFO-BT] Computer Science [cs]/Biotechnology, [CHIM.OTHE] Chemical Sciences/Other, Polymer adhesive, HCV, [SDV.IMM]Life Sciences [q-bio]/Immunology, Colorimetry, Adhesive, alpha-Fetoproteins, 0210 nano-technology, [CHIM.OTHE]Chemical Sciences/Other, Antibodies, Immobilized, Biotechnology, HeLa Cells
Abstract: http://dx.doi.org/10.1016/j.bios.2012.06.024; International audience; The present report describes the integration and application possibilities of a new microarray concept based on adhesive surface. The method was shown to enable the straightforward production of 384 and 1536-well plates modified with 100 and 25 spots per well, respectively. Such in-well densities were only possible thanks to the fabrication process which implies first the deposition of the microarray on a flat adhesive surface and then its assembly with bottomless 384 or 1536-well plates. The concept was also confronted to various applications such as oligonucleotide detection, localised cell culture onto spotted adhesion proteins and immobilisation of peptide or active antibodies for immunoassays. In the particular case of immunotesting, the study focused on liver diseases diagnosis and more particularly on the detection of either one liver cancer marker, the alpha-fetoprotein, or the detection of Hepatitis C Virus infection. In every cases, interesting performances were obtained directly in crude patient serum, proof of the robust and generic aspect of the platform.
Published: 2013

22. Human endogenous retrovirus protein activates innate immunity and promotes experimental allergic encephalomyelitis in mice

Author: Reza Firouzi, Evelyne Jouvin-Marche, Jacques Portoukalian, Patrice N. Marche, Suzana Marusic, Christian L. Villiers, Iuliana Popa, Hervé Perron, Jean-Louis Touraine, Christina Lomparski, Hei-Lanne Dougier-Reynaud, Jean-Baptiste Bertrand, Université de Lyon, GeNeuro, GeNeuro SA, Geneuro-Innovation, INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-ImmunAlp, ImmunAlp-ImmunAlp, Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Physico-chimie, pharmacotechnie, biopharmacie (PCPB), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Hook laboratories, Centre Hospitalier Lyon Sud [CHU - HCL] (CHLS), Hospices Civils de Lyon (HCL), Institutional support from INSERM/grants from Fondation CERA-Lyon and AFM-ARSEP, France., and Marche, Patrice
Subjects: Central Nervous System, Encephalomyelitis, Lipopolysaccharide Receptors, Gene Expression, Pregnancy Proteins, medicine.disease_cause, Autoimmunity, Mice, 0302 clinical medicine, Cells, Cultured, Mice, Knockout, 0303 health sciences, Multidisciplinary, Microglia, Recombinant Proteins, 3. Good health, medicine.anatomical_structure, [SDV.IMM]Life Sciences [q-bio]/Immunology, Medicine, Research Article, Encephalomyelitis, Autoimmune, Experimental, Multiple Sclerosis, [SDV.IMM] Life Sciences [q-bio]/Immunology, Science, Biology, Proinflammatory cytokine, Myelin oligodendrocyte glycoprotein, 03 medical and health sciences, Adjuvants, Immunologic, medicine, Animals, Humans, Neuroinflammation, 030304 developmental biology, Innate immune system, Gene Products, env, Dendritic Cells, medicine.disease, Immunity, Innate, Peptide Fragments, Mice, Inbred C57BL, Toll-Like Receptor 4, Disease Models, Animal, Immunology, TLR4, biology.protein, Myelin-Oligodendrocyte Glycoprotein, 030217 neurology & neurosurgery
Abstract: International audience; Multiple sclerosis (MS) is a complex multifactorial disease of the central nervous system (CNS) for which animal models have mainly addressed downstream immunopathology but not potential inducers of autoimmunity. In the absence of a pathogen known to cause neuroinflammation in MS, Mycobacterial lysate is commonly used in the form of complete Freund's adjuvant to induce autoimmunity to myelin proteins in Experimental Allergic Encephalomyelitis (EAE), an animal model for MS. The present study demonstrates that a protein from the human endogenous retrovirus HERV-W family (MSRV-Env) can be used instead of mycobacterial lysate to induce autoimmunity and EAE in mice injected with MOG, with typical anti-myelin response and CNS lesions normally seen in this model. MSRV-Env was shown to induce proinflammatory response in human macrophage cells through TLR4 activation pathway. The present results demonstrate a similar activation of murine dendritic cells and show the ability of MSRV-Env to trigger EAE in mice. In previous studies, MSRV-Env protein was reproducibly detected in MS brain lesions within microglia and perivascular macrophages. The present results are therefore likely to provide a model for MS, in which the upstream adjuvant triggering neuroinflammation is the one detected in MS active lesions. This model now allows pre-clinical studies with therapeutic agents targeting this endogenous retroviral protein in MS.
Published: 2013

23. Expression and Purification of a Soluble Functional form of the Platelet alphaIIbbeta3 Integrin

Author: Alain Duperray, Elisabeth Delachanal, Monica Alemany, Pascal Martinez, Gérard Marguerie, Danielle Gulino, Evelyne Concord, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Groupe de Recherche Sur Le Cancer du Poumon : Bases Moléculaires de la Progression Tumorale, Dépistage et Thérapie Génique, Institut Albert Bonniot-Institut National de la Santé et de la Recherche Médicale (INSERM), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche Cardiovasculaire de Lariboisiere, Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hémostase et thrombose, and Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Integrins, DNA, Complementary, Immunoprecipitation, Protein subunit, Molecular Sequence Data, Integrin, Platelet Glycoprotein GPIIb-IIIa Complex, Ligands, Biochemistry, Chromatography, Affinity, Cell Line, 03 medical and health sciences, 0302 clinical medicine, Native state, Humans, Amino Acid Sequence, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, COS cells, Base Sequence, biology, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, Precipitin Tests, Transmembrane protein, Solubility, Membrane protein, chemistry, 030220 oncology & carcinogenesis, biology.protein, Glycoprotein, Protein Binding
Abstract: Platelet glycoproteins alpha IIb and beta 3 are membrane proteins that associate to form a Ca(2+)-dependent heterodimer which constitutes an inducible member of the integrin family at the surface of the cell. To produce a soluble form of this complex, alpha IIb and beta 3 were both deleted of their transmembrane and cytoplasmic domains and were expressed in COS cells. Production of the truncated subunits and their mode of assembly were examined by immunoprecipitation experiments and compared to those of wild-type alpha IIb beta 3. Synthesis and processing of the truncated heterodimer proceeded via a pathway similar to that observed for the wild-type alpha IIb beta 3 in COS cells or in human megakaryocytes. The truncated beta 3 subunit associated with the Pro-truncated form of the alpha IIb subunit. This precursor form was not secreted. After proteolytic cleavage of the Pro-truncated alpha IIb, the mature heterodimer was secreted into the culture supernatant. To quantify the molar ratio of the various secreted soluble forms, an immunocapture assay was designed. All secreted tr-alpha IIb subunits associated with tr-beta 3. In contrast, tr-beta 3 was produced and secreted in excess as the free form. Immunoreactivity of the wild-type and soluble truncated complexes was identical since all the monoclonal antibodies used reacted with surface-located epitopes on both complexes. This indicated that the soluble truncated heterodimer adopted a native conformation. To purify this soluble heterodimer, tr-alpha IIb beta 3-containing culture supernatant was adsorbed on an RGDW-affinity column and eluted with a solution of the free peptide RGDW. In the RGD-eluted material, the amount of each subunit was stoichiometric, suggesting that the complex was not disrupted during purification. The capacity of the wild-type and truncated RGD-eluted complexes to interact with soluble fibrinogen was compared using a solid-phase immunocapture assay. tr-alpha IIb beta 3 and platelet alpha IIb beta 3 exhibited similar fibrinogen-binding capacity. For both complexes, these interactions were mediated by RGD and gamma fibrinogen signals.
Published: 1995

24. Inverse problems for the determination of traction forces by cells on a substrate: a comparison of two methods

Author: Alain Duperray, Richard Michel, Claude Verdier, Jocelyn Etienne, Baptiste Bedessem, Valentina Peschetola, Davide Carlo Ambrosi, DYFCOM, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), MOX, MOX-Dipartimento di Matematica (MOX), Politecnico di Milano [Milan] (POLIMI)-Politecnico di Milano [Milan] (POLIMI), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), and Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: Materials science, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Traction (engineering), Biomedical Engineering, Bioengineering, migration, 01 natural sciences, Zero force, 03 medical and health sciences, symbols.namesake, Cell Movement, Cell Line, Tumor, Humans, 0101 mathematics, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], 030304 developmental biology, Cell invasion, 0303 health sciences, Fourier Analysis, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], General Medicine, Mechanics, Inverse problem, Traction forces, eye diseases, Computer Science Applications, 010101 applied mathematics, Human-Computer Interaction, Stress field, regularization, Fourier transform, Urinary Bladder Neoplasms, Fourier analysis, displacement field, Displacement field, symbols
Abstract: International audience; Traction forces exerted by cells on soft elastic substrates are important for characterizing the types of mechanisms used for cell migration. Classical tools for the determination of traction forces include the knowledge of the displacement field thanks to fluorescent beads embedded into the substrate. Then, from the discrete beads displacements, an inverse problem is solved to obtain the stress field. Two currently used methods in the literature are the well-known Fourier Transform Traction Cytometry (FTTC) method and the adjoint method (AM), which are compared here. A real case is presented where the displacement field is known from cancer cell migration study. The two methods allow the recovery of the traction stresses and their results are compared. Similar results are seen as long as an adequate projection technique is used (zero force imposed outside the cell). It is found that the AM allows a finer resolution of the traction forces, in particular at the cell edge. This is a strong incentive to use this method for the investigation of cancer cell migration on soft substrates.
Published: 2012

25. Ytterbium-based bioprobes for near-infrared two-photon scanning laser microscopy imaging

Author: Adrien Bourdolle, Olivier Maury, Teddy Fauquier, Patrice L. Baldeck, Alexei Grichine, Chantal Andraud, Anthony D'Aléo, Sophie Brasselet, Sophie Brustlein, Alain Duperray, Laboratoire de Chimie - UMR5182 (LC), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École normale supérieure - Lyon (ENS Lyon)-Institut de Chimie du CNRS (INC), Laboratoire de Chimie, Catalyse, Polymères et Procédés, R 5265 (C2P2), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École supérieure de Chimie Physique Electronique de Lyon (CPE)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), MOSAIC (MOSAIC), Institut FRESNEL (FRESNEL), Centre National de la Recherche Scientifique (CNRS)-École Centrale de Marseille (ECM)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-École Centrale de Marseille (ECM)-Aix Marseille Université (AMU), Institut de Génomique Fonctionnelle de Lyon (IGFL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-École normale supérieure - Lyon (ENS Lyon), 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), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Lyon-Université de Lyon-École Supérieure de Chimie Physique Électronique de Lyon (CPE)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-École Centrale de Marseille (ECM)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Groupe de Recherche Sur Le Cancer du Poumon : Bases Moléculaires de la Progression Tumorale, Dépistage et Thérapie Génique, Institut Albert Bonniot-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École supérieure de Chimie Physique Electronique de Lyon (CPE)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Lyon (ENS Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Duperray, Alain, and Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)
Subjects: Ytterbium, Laser Microscopy, Materials science, Analytical chemistry, chemistry.chemical_element, Biosensing Techniques, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, 010402 general chemistry, 01 natural sciences, Catalysis, Two-photon excitation microscopy, Coordination Complexes, Cell Line, Tumor, Humans, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, ComputingMilieux_MISCELLANEOUS, NIR-to-NIR configuration, Microscopy, Microscopy, Confocal, Spectroscopy, Near-Infrared, 010405 organic chemistry, business.industry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Near-infrared spectroscopy, Biphotonic microscopy, Ytterbium complex, General Chemistry, General Medicine, [CHIM.ORGA] Chemical Sciences/Organic chemistry, Bioimaging, 0104 chemical sciences, chemistry, Lanthanide complex, [SPI.OPTI]Engineering Sciences [physics]/Optics / Photonic, Optoelectronics, business
Abstract: International audience; n.a
Published: 2012

26. Dynamic interplay between human myoblasts and 3D fibrin-based matrix

Author: Chiron, Stéphane, Tomczak, Carole, Duperray, Alain, Laine, Jeanne, Bonne, Gisele, Eder, Alexandra, Hansen, Arne, Eschenhagen, Thomas, Verdier, Claude, Coirault, Catherine, Thérapie des maladies du muscle strié, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Myologie, Université Pierre et Marie Curie - Paris 6 (UPMC)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Universitaetsklinikum Hamburg-Eppendorf = University Medical Center Hamburg-Eppendorf [Hamburg] (UKE), DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Verdier, Claude, Département de physiologie, Université Pierre et Marie Curie - Paris 6 (UPMC)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Thérapie des maladies du muscle strié, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre de référence des maladies rares neuromusculaires [CHU Pitié-Salpétriêre], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Service de Biochimie Métabolique [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre de référence des maladies rares neuromusculaires, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-CHU Pitié-Salpêtrière [AP-HP], DYFCOM, and Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: [PHYS.PHYS.PHYS-BIO-PH] Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], myotubes, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], myoblasts, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Fibrin gels, [SDV.BC.IC] Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB]
Abstract: International audience; Anchorage of muscle cells to the extracellular matrix is crucial for a range of fundamental biological processes including migration, survival and differentiation. Three-dimensional (3D) culture has been proposed to provide a more physiological in vitro model of muscle growth and differentiation than routine 2D cultures. However, muscle cell adhesion and cell-matrix interplay of engineered muscle tissue remain to be determined. We have characterized cell-matrix interactions in 3D muscle culture and analyzed their consequences on cell differentiation. Human myoblasts were embedded in a fibrin matrix cast between two posts, cultured until confluence, and then induced to differentiate. Myoblasts in 3D aligned along the longitudinal axis of the gel. They displayed actin stress fibers evenly distributed around the nucleus and a cortical mesh of thin actin filaments. Adhesion sites in 3D were smaller in size than in rigid 2D culture but expression of adhesion site proteins, including alpha5 integrin and vinculin, was higher in 3D compared with 2D (p,0.05). Myoblasts and myotubes in 3D exhibited thicker and ellipsoid nuclei instead of the thin disk-like shape of the nuclei in 2D (p,0.001). Differentiation kinetics were faster in 3D as demonstrated by higher mRNA concentrations of alpha-actinin and myosin. More important, the elastic modulus of engineered muscle tissues increased significantly from 3.5+/-0.8 to 7.4+/-4.7 kPa during proliferation (p,0.05) and reached 12.2+/-6.0 kPa during differentiation (p,0.05), thus attesting the increase of matrix stiffness during proliferation and differentiation of the myocytes. In conclusion, we reported modulations of the adhesion complexes, the actin cytoskeleton and nuclear shape in 3D compared with routine 2D muscle culture. These findings point to complex interactions between muscle cells and the surrounding matrix with dynamic regulation of the cell-matrix stiffness.
Published: 2012

27. Traction forces of cancer cells

Author: Davide Carlo Ambrosi, Alain Duperray, Valérie M. Laurent, Luigi Preziosi, Valentina Peschetola, Claude Verdier, Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Dipartimento di Matematica 'Giuseppe Peano' [Torino], Università degli studi di Torino (UNITO), MOX-Dipartimento di Matematica (MOX), Politecnico di Milano [Milan] (POLIMI), This work was partly funded by the European Commission (EC) through a Marie Curie Research Training Network (MRTN-CT-2004-503661) entitled 'Modeling, mathematical methods and computer simulation of tumor growth and therapy'., INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), DYFCOM, Politecnico di Torino Matematica, Politecnico di Milano MOX, European Project, Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Università degli studi di Torino = University of Turin (UNITO), Verdier, Claude, and Marie Curie Research Training Network MRTN-CT-2004-503661 - INCOMING
Subjects: 0206 medical engineering, Integrin, forces, Biomedical Engineering, Bioengineering, 02 engineering and technology, Biology, Focal adhesion, 03 medical and health sciences, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Myosin, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Cytoskeleton, Cell adhesion, Migration, 030304 developmental biology, 0303 health sciences, Cell adhesion molecule, [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Cell migration, General Medicine, 020601 biomedical engineering, Computer Science Applications, Cell biology, Human-Computer Interaction, adhesion, motility, Cancer cell, biology.protein, integrins, cancer cells
Abstract: Actes du congrès de la société de Biomécanique; International audience; Cell motility of cancer cells is a fundamental problem, and requires precise correlation between cell adhesive and rheological properties. The migration of cancer cells is studied in two dimensions, as cells are seeded onto functionnalized polyacrylamide gels. They migrate by developping focal adhesion sites and modulate their cytoskeleton dynamics by regulating actin and myosin. This research aims at understanding the forces developped by different cancer cells of moderate to high invasiveness in order to investigate their contractility as well as the adhesion molecules necessary for cell migration.
Published: 2011

28. Fabrication of Bio-Functionalised Polypyrrole Nanoarrays for Bio-Molecular Recognition

Author: Andrea Valsesia, Patricia Lisboa, Pascal Colpo, François Rossi, Carine Brakha, Patrice N. Marche, Marie-Bernadette Villiers, JRC Institute for Health and Consumer Protection (IHCP), European Commission - Joint Research Centre [Ispra] (JRC), 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), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Plasmore S.r.l., Plasmonics for Molecular Recognition, Plasmore S.r.l., and Marche, Patrice
Subjects: Fabrication, Materials science, [SDV.IMM] Life Sciences [q-bio]/Immunology, [SPI.NANO] Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, SPR, Nanotechnology, 02 engineering and technology, 010402 general chemistry, Polypyrrole, 01 natural sciences, chemistry.chemical_compound, Molecular recognition, polypyrrole, Surface plasmon resonance, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, Silicon oxide, [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, Nanoscopic scale, chemistry.chemical_classification, Biomolecule, Building and Construction, 021001 nanoscience & nanotechnology, 0104 chemical sciences, [SDV.IB.BIO] Life Sciences [q-bio]/Bioengineering/Biomaterials, chemistry, [CHIM.OTHE] Chemical Sciences/Other, Colloidal lithography, Nanosphere lithography, nanoarrays, [SDV.IMM]Life Sciences [q-bio]/Immunology, 0210 nano-technology, [CHIM.OTHE]Chemical Sciences/Other, immunoreaction
Abstract: International audience; The present study demonstrates that nanosphere lithography and electro-polymerization can be successfully combined to produce bioactive protein nanoarrays. In particular, we describe a method to produce well-defined nanoarrays of polypyrrole functionalized with biomolecules. The nanoarrayed surfaces were fabricated on gold coated surface plasmon resonance prisms by first creating silicon oxide or polyethylene oxide nanotemplate using nanosphere lithography. The nanotemplate was subsequently used to grow bio-functionalized polypyrrole nanoarrays by electrocopolymerization. Atomic force microscopy analysis showed that the fabricated surfaces have a well-organized 2D hexagonal geometry with nanoscale dimensions. The biological activity of the bio-functionalized polypyrrole was assessed by surface plasmon resonance detection. The results showed that the immobilized biomolecules within the nanoarrayed polypyrrole films had the necessary bioactivity for successful molecular recognition. Moreover the detection signals normalized to the bioactive area were increased by a factor 5 as compared to non-structured bio-functionalized polypyrrole in the nanoarrayed surfaces using polyethylene oxide.
Published: 2011

29. Enhanced susceptibility of T lymphocytes to oxidative stress in the absence of the cellular prion protein

Author: Aude-Garcia, Catherine, Villiers, Christian, Candéias, Serge, Garrel, Catherine, Bertrand, Caroline, Collin, Véronique, Marche, Patrice, Jouvin-Marche, Evelyne, 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), 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), Département de biologie intégrée, CHU Grenoble-Hôpital Michallon, Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), GIS-Prion #A108, CEA ToxNuc program #13-10, and European Community Project Immunoprion FP7-013144
Subjects: MESH: Glutathione, MESH: Oxidative Stress, animal diseases, T lymphocytes, MESH: Reactive Oxygen Species, Cellular prion protein, nervous system diseases, Thymus, Redox balance, MESH: T-Lymphocytes, MESH: Prions, Oxidative stress, MESH: Gene Deletion, MESH: Mice, Inbred C57BL, MESH: Hydrogen Peroxide, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Animals, MESH: Mice, MESH: Cells, Cultured
Abstract: International audience; The cellular prion glycoprotein (PrP(C)) is ubiquitously expressed but its physiologic functions remain enigmatic, particularly in the immune system. Here, we demonstrate in vitro and in vivo that PrP(C) is involved in T lymphocytes response to oxidative stress. By monitoring the intracellular level of reduced glutathione, we show that PrP(-/-) thymocytes display a higher susceptibility to H(2)O(2) exposure than PrP(+/+) cells. Furthermore, we find that in mice fed with a restricted diet, a regimen known to increase the intracellular level of ROS, PrP(-/-) thymocytes are more sensitive to oxidative stress. PrP(C) function appears to be specific for oxidative stress, since no significant differences are observed between PrP(-/-) and PrP(+/+) mice exposed to other kinds of stress. We also show a marked evolution of the redox status of T cells throughout differentiation in the thymus. Taken together, our results clearly ascribe to PrP(C) a protective function in thymocytes against oxidative stress.
Published: 2011

30. On chip real time monitoring of B-cells hybridoma secretion of immunoglobulin

Author: Patrice N. Marche, Sarah Milgram, Sandra Cortès, Marie-Bernadette Villiers, Yoann Roupioz, Arnaud Buhot, Thierry Livache, Structures et propriétés d'architectures moléculaire (SPRAM - UMR 5819), Institut Nanosciences et Cryogénie (INAC), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), and Marche, Patrice
Subjects: MESH: Immunoglobulins, Cell Culture Techniques, MESH: Equipment Failure Analysis, Biosensing Techniques, 01 natural sciences, Real-time detection, Electrochemistry, Biochip, Immunoassay, 0303 health sciences, B-Lymphocytes, MESH: Protein Array Analysis, General Medicine, Equipment Design, Protein micro-array, MESH: Surface Plasmon Resonance, medicine.anatomical_structure, Biochemistry, [SDV.IMM]Life Sciences [q-bio]/Immunology, Antibody, MESH: Immunoassay, MESH: Biosensing Techniques, Biotechnology, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Hybridomas, Biomedical Engineering, Biophysics, Protein Array Analysis, Immunoglobulins, Biology, Cell Line, Surface plasmon resonance imaging, 03 medical and health sciences, Immune system, Antigen, MESH: B-Lymphocytes, medicine, Humans, Secretion, B cell, 030304 developmental biology, MESH: Cell Culture Techniques, Hybridomas, MESH: Humans, Blood cell, 010401 analytical chemistry, Cellular secretion, Surface Plasmon Resonance, Molecular biology, In vitro, 0104 chemical sciences, MESH: Cell Line, Equipment Failure Analysis, biology.protein, Biosensor, MESH: Equipment Design
Abstract: International audience; The secretions of molecules by cells are of tremendous interest for both fundamental insights studies and medical purposes. In this study, we propose a new biochip-based approach for the instantaneous monitoring of protein secretions, using antibody production by B lymphocytes cultured in vitro. This was possible thanks to the Surface Plasmon Resonance imaging (SPRi) of a protein biochip where antigen proteins (Hen Egg Lysozyme, HEL) were micro-arrayed along with series of control proteins. B cell hybridomas were cultured on the chip and the secretion of immunoglobulins (antibody) specific to HEL was monitored in real-time and detected within only few minutes rather than after a 30-60 min incubation with standard ELISA experiments. This fast and sensitive detection was possible thanks to the sedimentation of the cells on the biochip sensitive surface, where local antibody concentrations are much higher before dilution in the bulk medium. An other interesting feature of this approach for the secretion monitoring was the independence of the SPR response--after normalization--regarding to the density of the surface-immobilized probes. Such biosensor might thus pave the way to new tools capable of both qualitative and semi-quantitative analysis of proteins secreted by other immune cells.
Published: 2011

31. Initial Dynamics of Cell Spreading Are Governed by Dissipation in the Actin Cortex

Author: Alain Duperray, Jocelyn Etienne, Etienne, Jocelyn, DYnamique des Fluides COmplexes et Morphogénèse [Grenoble] (DYFCOM), Laboratoire Interdisciplinaire de Physique [Saint Martin d’Hères] (LIPhy), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Experiments were performed at the microscopy facility of the Institut Albert Bonniot. This equipment was partly funded by the Association pour la Recherche sur le Cancer (Villejuif, France) and the Nanobio program., DYFCOM, Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), and Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)
Subjects: [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Finite elements, Biophysics, 01 natural sciences, Models, Biological, Modelling, 03 medical and health sciences, Cell Line, Tumor, 0103 physical sciences, Cell cortex, Humans, Cellular Biophysics and Electrophysiology, 010306 general physics, Cell adhesion, Cytoskeleton, Cell Shape, Actin, 030304 developmental biology, 0303 health sciences, Deformation (mechanics), [PHYS.PHYS.PHYS-BIO-PH] Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Chemistry, Viscosity, Adhesion, Actomyosin, Dissipation, Actins, Cell biology, Biomechanical Phenomena, Spectrometry, Fluorescence, Dissipative system, Cell biophysics
Abstract: 11 pages; International audience; The initial stages of spreading of a suspended cell onto a substrate under the effect of (specific or nonspecific) adhesion exhibit a universal behavior, which is cell-type independent. We show that this behavior is governed only by cell-scale phenomena. This can be understood if the main retarding force that opposes cell adhesion is of mechanical origin, that is, dissipation occurring during the spreading. By comparing several naive models that generate different patterns of dissipation, we show by numerical simulation that only dissipation due to the deformation of the actin cortex is compatible with the experimental observations. This viscous-like dissipation corresponds to the energetic cost of rearranging the cytoskeleton, and is the trace of all dissipative events occurring in the cell cortex during the early spreading, such as the binding and unbinding of cross-linkers and molecular friction.
Published: 2011

32. Enhanced susceptibility of T lymphocytes to oxidative stress in the absence of the cellular prion protein.: PrPC and oxidative stress in T lymphocytes

Author: Aude-Garcia, Catherine, Villiers, Christian, Candéias, Serge, Garrel, Catherine, Bertrand, Caroline, Collin, Véronique, Marche, Patrice, Jouvin-Marche, Evelyne, Aude-Garcia, Catherine, 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), 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), Département de biologie intégrée, CHU Grenoble-Hôpital Michallon, Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), GIS-Prion #A108, CEA ToxNuc program #13-10, and European Community Project Immunoprion FP7-013144
Subjects: MESH: Glutathione, MESH: Oxidative Stress, [SDV.IMM] Life Sciences [q-bio]/Immunology, animal diseases, T lymphocytes, MESH: Reactive Oxygen Species, Cellular prion protein, nervous system diseases, Thymus, Redox balance, MESH: T-Lymphocytes, MESH: Prions, Oxidative stress, MESH: Gene Deletion, MESH: Mice, Inbred C57BL, MESH: Hydrogen Peroxide, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Animals, MESH: Mice, MESH: Cells, Cultured
Abstract: International audience; The cellular prion glycoprotein (PrP(C)) is ubiquitously expressed but its physiologic functions remain enigmatic, particularly in the immune system. Here, we demonstrate in vitro and in vivo that PrP(C) is involved in T lymphocytes response to oxidative stress. By monitoring the intracellular level of reduced glutathione, we show that PrP(-/-) thymocytes display a higher susceptibility to H(2)O(2) exposure than PrP(+/+) cells. Furthermore, we find that in mice fed with a restricted diet, a regimen known to increase the intracellular level of ROS, PrP(-/-) thymocytes are more sensitive to oxidative stress. PrP(C) function appears to be specific for oxidative stress, since no significant differences are observed between PrP(-/-) and PrP(+/+) mice exposed to other kinds of stress. We also show a marked evolution of the redox status of T cells throughout differentiation in the thymus. Taken together, our results clearly ascribe to PrP(C) a protective function in thymocytes against oxidative stress.
Published: 2011

33. Peptide-protein microarrays and surface plasmon resonance detection: biosensors for versatile biomolecular interaction analysis

Author: Thierry Livache, Sandra Cortès, Jean-Pierre Lavergne, Paul Dény, Patrice N. Marche, Christophe A. Marquette, Carine Brakha, Marie-Bernadette Villiers, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), 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), Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), Physiopathologie moléculaire et nouveaux traitements des hépatites virales, Université de Lyon-Université de Lyon-IFR62-Institut National de la Santé et de la Recherche Médicale (INSERM), Structures et propriétés d'architectures moléculaire (SPRAM - UMR 5819), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Lyon-Université de Lyon-École Supérieure de Chimie Physique Électronique de Lyon (CPE)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), and Marche, Patrice
Subjects: [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Microarray, Analytical chemistry, Peptide, Biosensing Techniques, Ligands, 01 natural sciences, Mice, Electrochemistry, MESH: Ligands, MESH: Animals, Surface plasmon resonance, [INFO.INFO-BT]Computer Science [cs]/Biotechnology, chemistry.chemical_classification, 0303 health sciences, Chemistry, MESH: Protein Array Analysis, Surface plasmon, General Medicine, peptide, MESH: Surface Plasmon Resonance, Metals, Protein microarray, [SDV.IMM]Life Sciences [q-bio]/Immunology, DNA microarray, MESH: Immobilized Proteins, MESH: Biosensing Techniques, Biotechnology, Protein Binding, MESH: Ions, [SDV.IMM] Life Sciences [q-bio]/Immunology, [SPI.NANO] Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, Biomedical Engineering, Biophysics, Protein Array Analysis, Nanotechnology, Antibodies, Cell Line, 03 medical and health sciences, Ion binding, Animals, Humans, MESH: Protein Binding, Surface Plasmon Resonance imaging, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, MESH: Mice, 030304 developmental biology, Ions, MESH: Humans, MESH: Metals, MESH: Antibodies, 010401 analytical chemistry, Surface Plasmon Resonance, cell, 0104 chemical sciences, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, MESH: Cell Line, Immobilized Proteins, [INFO.INFO-BT] Computer Science [cs]/Biotechnology, ion, protein, Biosensor
Abstract: International audience; Biosensors in microarray format provide promising tools for high-throughput analyses of complex samples. Although they are able to detect, quantify and characterize a multitude of compounds, most of the available devices are specialized in the analysis of one type of interaction, limiting their application to a define area. The aim of our work was to develop and characterize versatile protein (or peptide) microarrays suitable for the simultaneous analysis of a large panel of biological interactions. Our system involved a simple procedure to immobilized proteins or peptides, based on pyrrole electropolymerization, and ligand binding was detected by imaging the surface plasmon resonance. We demonstrated its suitability in three different contexts, i.e. humoral response characterization, ion binding analysis and cell detection. This work evidences the potentiality of this approach which allows multiparametric, high-throughput and label-free analysis of biological samples suitable for the detection of compounds as various as proteins, ions or cells and the characterization of their interaction with peptides or proteins.
Published: 2010

34. Breakdown of cell-collagen networks through collagen remodeling

Author: Anaïs Gérard, Claude Verdier, Alain Duperray, Andreea Iordan, Alexei Grichine, Laboratoire de Spectrométrie Physique (LSP), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), 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)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Marie Curie Research Training Network (MRTN-CT-2004-503661), and Verdier, Claude
Subjects: collagen, reflectance, Physiology, Confocal, Cell, CHO Cells, law.invention, Cricetulus, Cell Movement, Confocal microscopy, law, Cricetinae, Physiology (medical), [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Collagen network, Cell Adhesion, medicine, Animals, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Cell Shape, remodeling, Microscopy, Confocal, Chemistry, [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], tissue, Anatomy, Reflectivity, Elasticity, Extracellular Matrix, medicine.anatomical_structure, confocal, Biophysics, rheology, Stress, Mechanical, Gels
Abstract: International audience; Collagen model tissues are analyzed, which consist of cells embedded in a collagen matrix at different concentrations (of cells and collagen). Rheological properties are measured and complementary confocal microscopy analyses are carried out. An important feature is observed, corresponding to the breakdown of the collagen network (i.e. decrease in network elasticity) for high collagen concentrations, due to the presence of cells. Thanks to confocal microscopy, we show that cells elongate within the gel and can remodel it, this being a concentration-dependent feature. A careful analysis of the remodelling process shows that cells can attract collagen in their close neighborhood, this being an irreversible process, and that migrating cells create collagen-depleted regions behind them.
Published: 2010

35. Characterization and role of intra-hepatic regulatory T cells in chronic hepatitis C pathogenesis.: Regulatory T cells in chronic hepatitis C

Author: Sturm, Nathalie, Thélu, Marie-Ange, Camous, Xavier, Dimitrov, Guéorgui, Ramzan, Muhammad, Dufeu-Duchesne, Tania, Bonorino, Paula, Guillermet, Christiane, Brambilla, Elisabeth, Arvers, Philippe, Pernollet, Martine, Leroy, Vincent, Zarski, Jean-Pierre, Marche, Patrice, Jouvin-Marche, Evelyne, 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), Département d'anatomie et cythologie pathologique, CHU Grenoble-Hôpital Michallon, INSERM U823, équipe 2 (Bases Moléculaires de la Progression des Cancers du Poumon), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département d'anatomie et cythologie pathologique, CHU Grenoble-Hôpital Michallon-Hôpital Michallon, Centre de Recherches du Service de Santé des Armées (CRSSA), Service de Santé des Armées, INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by the institutional Grants from Institut National de la Santé et de la Recherche Médicale (INSERM), by a specific grant from the Agence Nationale pour la Recherche sur le Sida (ANRS), by specific support from Centre National de la Recherche Scientifique (CNRS), and in part by the 'Biotherapic' and 'AlphaVac' programs of the 'Pôle de Compétitivité Lyon Biopôle'. M.R. was supported by a fellowship from Higher Education Commission (HEC) of Pakistan, X.C. was supported by a fellowship from Région-Rhône Alpes actions in the 'Clusters 10', and P.B. was supported by fellowships from French government and ANRS.
Subjects: Liver, Hepatitis C Virus (HCV), FoxP3, T lymphocytes, [SDV.IMM]Life Sciences [q-bio]/Immunology, hemic and immune systems, chemical and pharmacologic phenomena, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Regulatory T cells, Immunohistochemistry
Abstract: International audience; BACKGROUND & AIMS: In chronic hepatitis C (CHC), HCV-specific T-cell responses are often dysfunctionnal. In vitro data point out that regulatory T cells (Treg) are able to suppress HCV-specific lymphocyte proliferation and cytokine secretion but their implication in this pathology is still debated. METHODS: Three complementary approaches were performed to investigate phenotype, frequency or localization of intra-hepatic Treg in treatment naïve CHC patients. Double immunohistochemical analysis was performed in 20 formalin-fixed biopsies with CD8/FoxP3 and CD4/FoxP3 antibodies. Cellular markers and cytokines were investigated by quantitative RT-PCR in 27 additional frozen biopsies. Eight other fresh liver biopsies were selected for complementary analysis of immunophenotyping and frequency of intra-hepatic Treg. RESULTS: Immunohistochemical analyses showed the presence of intra-hepatic CD4(+)FoxP3(+)T cells while CD8(+)FoxP3(+)T cells were very scarce. CD4(+)FoxP3(+)T cells were located in necro-inflammatory areas in contact with CD8(+)T cells, suggesting that Treg-mediated inhibition of CD8(+)T cell proliferation may occur by cell-cell contact. RT-PCR analyses showed strong correlations between CD8, FoxP3, and IL-10 with emergence of four distinct gene clusters, CD8-FoxP3, CD8-IL-10, TGF-beta-IL-10, and TNF-alpha-TGF-beta. No correlation was found between serum viral load and any immune markers. Interestingly, the FoxP3(+)/CD8(+) cells ratio significantly decreased in severe fibrosis (F>3) due to the dramatic decline of FoxP3 cells. CONCLUSIONS: This study provides new insights into the histological localization of Treg within HCV-infected liver, with a special accumulation of CD4(+)FoxP3(+)Treg cells in necro-inflammatory areas, in contact with CD8(+)T cells. Our results suggest a link between Treg, CD8, and IL-10 which altogether could balance immune responses against the virus to avoid immunopathogenesis.
Published: 2010

36. Characterization and role of intra-hepatic regulatory T cells in chronic hepatitis C pathogenesis.: Regulatory T cells in chronic hepatitis C

Author: Sturm, Nathalie, Thélu, Marie-Ange, Camous, Xavier, Dimitrov, Guéorgui, Ramzan, Muhammad, Dufeu-Duchesne, Tania, Bonorino, Paula, Guillermet, Christiane, Brambilla, Elisabeth, Arvers, Philippe, Pernollet, Martine, Leroy, Vincent, Zarski, Jean-Pierre, Marche, Patrice, Jouvin-Marche, Evelyne, Marche, Patrice, 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), Département d'anatomie et cythologie pathologique, CHU Grenoble-Hôpital Michallon, INSERM U823, équipe 2 (Bases Moléculaires de la Progression des Cancers du Poumon), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département d'anatomie et cythologie pathologique, CHU Grenoble-Hôpital Michallon-Hôpital Michallon, Centre de Recherches du Service de Santé des Armées (CRSSA), Service de Santé des Armées, INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by the institutional Grants from Institut National de la Santé et de la Recherche Médicale (INSERM), by a specific grant from the Agence Nationale pour la Recherche sur le Sida (ANRS), by specific support from Centre National de la Recherche Scientifique (CNRS), and in part by the 'Biotherapic' and 'AlphaVac' programs of the 'Pôle de Compétitivité Lyon Biopôle'. M.R. was supported by a fellowship from Higher Education Commission (HEC) of Pakistan, X.C. was supported by a fellowship from Région-Rhône Alpes actions in the 'Clusters 10', and P.B. was supported by fellowships from French government and ANRS.
Subjects: Liver, [SDV.IMM] Life Sciences [q-bio]/Immunology, Hepatitis C Virus (HCV), FoxP3, T lymphocytes, [SDV.IMM]Life Sciences [q-bio]/Immunology, hemic and immune systems, chemical and pharmacologic phenomena, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Regulatory T cells, Immunohistochemistry, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology
Abstract: International audience; BACKGROUND & AIMS: In chronic hepatitis C (CHC), HCV-specific T-cell responses are often dysfunctionnal. In vitro data point out that regulatory T cells (Treg) are able to suppress HCV-specific lymphocyte proliferation and cytokine secretion but their implication in this pathology is still debated. METHODS: Three complementary approaches were performed to investigate phenotype, frequency or localization of intra-hepatic Treg in treatment naïve CHC patients. Double immunohistochemical analysis was performed in 20 formalin-fixed biopsies with CD8/FoxP3 and CD4/FoxP3 antibodies. Cellular markers and cytokines were investigated by quantitative RT-PCR in 27 additional frozen biopsies. Eight other fresh liver biopsies were selected for complementary analysis of immunophenotyping and frequency of intra-hepatic Treg. RESULTS: Immunohistochemical analyses showed the presence of intra-hepatic CD4(+)FoxP3(+)T cells while CD8(+)FoxP3(+)T cells were very scarce. CD4(+)FoxP3(+)T cells were located in necro-inflammatory areas in contact with CD8(+)T cells, suggesting that Treg-mediated inhibition of CD8(+)T cell proliferation may occur by cell-cell contact. RT-PCR analyses showed strong correlations between CD8, FoxP3, and IL-10 with emergence of four distinct gene clusters, CD8-FoxP3, CD8-IL-10, TGF-beta-IL-10, and TNF-alpha-TGF-beta. No correlation was found between serum viral load and any immune markers. Interestingly, the FoxP3(+)/CD8(+) cells ratio significantly decreased in severe fibrosis (F>3) due to the dramatic decline of FoxP3 cells. CONCLUSIONS: This study provides new insights into the histological localization of Treg within HCV-infected liver, with a special accumulation of CD4(+)FoxP3(+)Treg cells in necro-inflammatory areas, in contact with CD8(+)T cells. Our results suggest a link between Treg, CD8, and IL-10 which altogether could balance immune responses against the virus to avoid immunopathogenesis.
Published: 2010

37. Hepatitis C Virus (HCV) Evades NKG2D-Dependent NK Cell Responses through NS5A-Mediated Imbalance of Inflammatory Cytokines

Author: Sène, Damien, Levasseur, Franck, Abel, Michal, Lambert, Marion, Camous, Xavier, Hernandez, Céline, Pène, Véronique, Rosenberg, Arielle, Jouvin-Marche, Evelyne, Marche, Patrice, Cacoub, Patrice, Caillat-Zucman, Sophie, Marche, Patrice, Immunologie et génétique du diabète de type 1, génétique multifactorielle en endocrinologie pédiatrique (U986), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de médecine interne [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), 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), Virologie de l'hépatite C (EA 4474), Université Paris Descartes - Paris 5 (UPD5), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), and This work was supported by INSERM AVENIR program to S. C-Z. D. Sène received a grant from Association pour la Recherche contre le Cancer (ARC). F. Levasseur was supported by the French Ministry of Research. X. Camoux was supported by a fellowship from the 'Cluster 10: Infectiology' of the Rhone-Alpes Region, France.
Subjects: [SDV.IMM] Life Sciences [q-bio]/Immunology, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, [SDV.IMM]Life Sciences [q-bio]/Immunology, chemical and pharmacologic phenomena
Abstract: International audience; Understanding how hepatitis C virus (HCV) induces and circumvents the host's natural killer (NK) cell-mediated immunity is of critical importance in efforts to design effective therapeutics. We report here the decreased expression of the NKG2D activating receptor as a novel strategy adopted by HCV to evade NK-cell mediated responses. We show that chronic HCV infection is associated with expression of ligands for NKG2D, the MHC class I-related Chain (MIC) molecules, on hepatocytes. However, NKG2D expression is downmodulated on circulating NK cells, and consequently NK cell-mediated cytotoxic capacity and interferon-γ production are impaired. Using an endotoxin-free recombinant NS5A protein, we show that NS5A stimulation of monocytes through Toll-like Receptor 4 (TLR4) promotes p38- and PI3 kinase-dependent IL-10 production, while inhibiting IL-12 production. In turn, IL-10 triggers secretion of TGFβ which downmodulates NKG2D expression on NK cells, leading to their impaired effector functions. Moreover, culture supernatants of HCV JFH1 replicating Huh-7.5.1 cells reproduce the effect of recombinant NS5A on NKG2D downmodulation. Exogenous IL-15 can antagonize the TGFβ effect and restore normal NKG2D expression on NK cells. We conclude that NKG2D-dependent NK cell functions are modulated during chronic HCV infection, and demonstrate that this alteration can be prevented by exogenous IL-15, which could represent a meaningful adjuvant for therapeutic intervention.
Published: 2010

38. Hepatitis C Virus (HCV) Evades NKG2D-Dependent NK Cell Responses through NS5A-Mediated Imbalance of Inflammatory Cytokines. : HCV evasion to NKG2D-mediated response

Author: Sène, Damien, Levasseur, Franck, Abel, Michal, Lambert, Marion, Camous, Xavier, Hernandez, Céline, Pène, Véronique, Rosenberg, Arielle, Jouvin-Marche, Evelyne, Marche, Patrice, Cacoub, Patrice, Caillat-Zucman, Sophie, Immunologie et génétique du diabète de type 1, génétique multifactorielle en endocrinologie pédiatrique (U986), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM), Service de médecine interne [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) - CHU Pitié-Salpêtrière [APHP], 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), Virologie de l'hépatite C (EA 4474), Université Paris Descartes - Paris 5 (UPD5), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF) - CHU Grenoble - EFS - Institut National de la Santé et de la Recherche Médicale (INSERM) - Université Joseph Fourier - Grenoble 1 (UJF) - CHU Grenoble - EFS - Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by INSERM AVENIR program to S. C-Z. D. Sène received a grant from Association pour la Recherche contre le Cancer (ARC). F. Levasseur was supported by the French Ministry of Research. X. Camoux was supported by a fellowship from the 'Cluster 10: Infectiology' of the Rhone-Alpes Region, France., Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), and Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: [SDV.IMM] Life Sciences [q-bio]/Immunology, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, [SDV.IMM]Life Sciences [q-bio]/Immunology, chemical and pharmacologic phenomena
Abstract: International audience; Understanding how hepatitis C virus (HCV) induces and circumvents the host's natural killer (NK) cell-mediated immunity is of critical importance in efforts to design effective therapeutics. We report here the decreased expression of the NKG2D activating receptor as a novel strategy adopted by HCV to evade NK-cell mediated responses. We show that chronic HCV infection is associated with expression of ligands for NKG2D, the MHC class I-related Chain (MIC) molecules, on hepatocytes. However, NKG2D expression is downmodulated on circulating NK cells, and consequently NK cell-mediated cytotoxic capacity and interferon-γ production are impaired. Using an endotoxin-free recombinant NS5A protein, we show that NS5A stimulation of monocytes through Toll-like Receptor 4 (TLR4) promotes p38- and PI3 kinase-dependent IL-10 production, while inhibiting IL-12 production. In turn, IL-10 triggers secretion of TGFβ which downmodulates NKG2D expression on NK cells, leading to their impaired effector functions. Moreover, culture supernatants of HCV JFH1 replicating Huh-7.5.1 cells reproduce the effect of recombinant NS5A on NKG2D downmodulation. Exogenous IL-15 can antagonize the TGFβ effect and restore normal NKG2D expression on NK cells. We conclude that NKG2D-dependent NK cell functions are modulated during chronic HCV infection, and demonstrate that this alteration can be prevented by exogenous IL-15, which could represent a meaningful adjuvant for therapeutic intervention.
Published: 2010

39. Tumor cell/endothelial cell tight contact upregulates endothelial adhesion molecule expression mediated by NFkappaB: differential role of the shear stress

Author: Roxana Chotard-Ghodsnia, Alain Duperray, Oualid Haddad, Claude Verdier, Duperray, Alain, 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), Science des Procédés Céramiques et de Traitements de Surface (SPCTS), Université de Limoges (UNILIM)-Ecole Nationale Supérieure de Céramique Industrielle (ENSCI)-Institut des Procédés Appliqués aux Matériaux (IPAM), Université de Limoges (UNILIM)-Université de Limoges (UNILIM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie Physique (LSP), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), and Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Cell signaling, Endothelium, Adhesion molecule, Interaction, Blotting, Western, Vascular Cell Adhesion Molecule-1, Cell Communication, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Flow cytometry, Metastasis, 03 medical and health sciences, 0302 clinical medicine, Cell Line, Tumor, medicine, Humans, Cell adhesion, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Cells, Cultured, 030304 developmental biology, Cancer, 0303 health sciences, medicine.diagnostic_test, Cell adhesion molecule, Flow, NF-kappa B, Endothelial Cells, Cell Biology, Flow Cytometry, Intercellular Adhesion Molecule-1, Coculture Techniques, Extravasation, Up-Regulation, Cell biology, Endothelial stem cell, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Stress, Mechanical, Signal transduction, E-Selectin, Signal Transduction, NFκB
Abstract: International audience; Cancer metastasis is a multistep process involving cell-cell interactions, but little is known about the adhesive interactions and signaling events during extravasation of tumor cells (TCs). In this study, cell adhesion molecule (CAM) expression was investigated using an in vitro assay, in which TCs were seeded onto an endothelial cell (ECs) monolayer and cocultured during 5 hours. Flow cytometry, confocal microscopy as well as western blot analysis indicated that endothelial ICAM-1 (Inter Cellular Adhesion Molecule-1), VCAM-1 (Vascular Adhesion Molecule-1) and E-selectin were up-regulated after TC-EC coculture, whereas no change was observed for CAMs expression in tumor cells. This increased CAMs expression required tight contact between TCs and ECs. Incubation of ECs with the pyrrolidine-dithiocarbamate NFkappaB inhibitor prior to coculture, fully prevented coculture-induced expression of endothelial CAMs. Using specific blocking antibodies we showed an implication of ICAM-1 and VCAM-1 for TCs extravasation and VCAM-1 for adhesion. Moreover, fluid flow experiments revealed that high shear stress totally abolished coculture-induced as well as TNFalpha-induced CAMs over-expression. This study suggests that TCs could act as a potent inflammatory stimulus on ECs by inducing CAMs expression via NFkappaB activation, and that this action can be modulated by shear stress.
Published: 2010

40. Analysis of the toxicity of gold nano particles on the immune system: effect on dendritic cell functions

Author: Marie-Bernadette Villiers, Rachel Couderc, Patrice N. Marche, Heidi Freitas, Christian L. Villiers, Marche, Patrice, 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), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), and CellNanoTox project (FP6-NMP-CT-2006-032731)
Subjects: Materials science, [SDV.IMM] Life Sciences [q-bio]/Immunology, medicine.medical_treatment, [SPI.NANO] Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, education, Endocytic cycle, chemical and pharmacologic phenomena, Bioengineering, Nanotechnology, 02 engineering and technology, Article, 03 medical and health sciences, Immune system, cytokine, medicine, General Materials Science, dendritic cells, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, Cytotoxicity, health care economics and organizations, 030304 developmental biology, 0303 health sciences, technology, industry, and agriculture, General Chemistry, Dendritic cell, biochemical phenomena, metabolism, and nutrition, gold, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Atomic and Molecular Physics, and Optics, Cell biology, [SDV.TOX] Life Sciences [q-bio]/Toxicology, immune system, medicine.anatomical_structure, Cytokine, Colloidal gold, Modeling and Simulation, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Nanoparticles, cytotoxicity, [SDV.IMM]Life Sciences [q-bio]/Immunology, Bone marrow, 0210 nano-technology, Intracellular
Abstract: International audience; The effect of manufactured gold nanoparticles (NP) on the immune system was analysed through their ability to perturb the functions of dendritic cells (DC), a major actor of both innate and acquired immune responses. For this purpose, DCs were produced in culture from mouse bone marrow progenitors.The analysis of the viability of the cells after their incubation in the presence of gold NP shows that these NP are not cytotoxics even at high concentration. Furthermore, the phenotype of the DC is unchanged after the addition of NP, indicating that there is no activation of the DC. But the analysis of the cells at the intracellular level reveals important amounts of gold NP amassing in endocytic compartments. Furthermore, the secretion of cytokines is significantly modified after such internalisation indicating a potential perturbation of the immune response.
Published: 2010

41. Review. Rheological properties of biological materials

Author: Verdier, Claude, Etienne, Jocelyn, Duperray, Alain, Preziosi, Luigi, Laboratoire de Spectrométrie Physique (LSP), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Dipartimento di Matematica 'Giuseppe Peano' [Torino], Università degli studi di Torino (UNITO), réseau européen RTN, Verdier, Claude, Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), and Università degli studi di Torino = University of Turin (UNITO)
Subjects: cell mechanics, viscoelastic, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], tissues, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Rheology
Abstract: International audience; Eucaryotic cells and biological materials are described from a rheological point of view. Single cell properties give rise to typical microrheological properties which can aect cell behaviour, in close connection with their adhesion properties. Single cell properties are also important in the context of multicellular systems, i.e. in biological tissues. Results from experiments are analyzed and models proposed both at the cellular scale and the macroscopic scale.
Published: 2009

42. A lymphotoxin-driven pathway to hepatocellular carcinoma

Author: Haybaeck, Johannes, Zeller, Nicolas, Wolf, Monika Julia, Weber, Achim, Wagner, Ulrich, Kurrer, Michael Odo, Bremer, Juliane, Lezzi, Giandomenica, Graf, Rolf, Clavien, Pierre-Alain, Thimme, Robert, Blum, Hubert, Nedospasov, Sergei A., Zatloukal, Kurt, Ramzan, Muhammad, Ciesek, Sandra, Pietschmann, Thomas, Marche, Patrice, Karin, Michael, Kopf, Manfred, Browning, Jeffrey, Aguzzi, Adriano, Heikenwalder, Mathias, Institute of Neuropathology, University hospital of Zurich [Zurich], Institutes of Neuropathology and Clinical Pathology, Functional Genomics Center Zurich, Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich)-Universität Zürich [Zürich] = University of Zurich (UZH), Department of Pathology, Aarau Catonal Hospital, Institute of Integrative Biology, Molecular Biomedicine, Swiss Hepato-Pancreatico-Biliary Center, Department of Internal Medicine, University of Freiburg [Freiburg], Engelhardt Institute of Molecular Biology (ISTC), Russian Academy of Sciences [Moscow] (RAS), Institute of Pathology, Medical University Graz, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Division of Experimental Virology, Centre for Experimental and Clinical Infection Research (TWINCORE), Helmholtz Centre for Infection Research (HZI)-Medizinische Hochschule Hannover (MHH)-Helmholtz Centre for Infection Research (HZI)-Medizinische Hochschule Hannover (MHH), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), University of California [San Diego] (UC San Diego), University of California, Department of Immunobiology, Biogen [Cambridge, USA], and Part of the project was achieved with support of the Agence Nationale pour la Recherche sur le Sida (ANRS) and from Pole de Compétitivité LyonBiopole, France. MR was supported by the Higher Education Commission of Pakistan.
Subjects: [SDV.IMM]Life Sciences [q-bio]/Immunology, [SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, neoplasms, digestive system diseases
Abstract: International audience; Hepatitis B and C viruses (HBV and HCV) cause chronic hepatitis and hepatocellular carcinoma (HCC) by poorly understood mechanisms. We show that cytokines lymphotoxin (LT) alpha and beta and their receptor (LTbetaR) are upregulated in HBV- or HCV-induced hepatitis and HCC. Liver-specific LTalphabeta expression in mice induces liver inflammation and HCC, causally linking hepatic LT overexpression to hepatitis and HCC. Development of HCC, composed in part of A6(+) oval cells, depends on lymphocytes and IKappa B kinase beta expressed by hepatocytes but is independent of TNFR1. In vivo LTbetaR stimulation implicates hepatocytes as the major LT-responsive liver cells, and LTbetaR inhibition in LTalphabeta-transgenic mice with hepatitis suppresses HCC formation. Thus, sustained LT signaling represents a pathway involved in hepatitis-induced HCC.
Published: 2009

43. The envelope of human endogenous retrovirus in neuro-inflammation

Author: Christina Lomparski, Hei-Lanne Dougier, Hervé Perron, Alain Duperray, Patrice N. Marche, Corinne Bernard, Christian L. Villiers, Evelyne Jouvin-Marche, 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), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Geneuro, GeNeuro, and BMC, Ed.
Subjects: lcsh:Immunologic diseases. Allergy, 0303 health sciences, business.industry, Human endogenous retrovirus, Bioinformatics, Virology, 3. Good health, Neuro inflammation, 03 medical and health sciences, 0302 clinical medicine, Infectious Diseases, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Poster Presentation, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, ComputingMethodologies_DOCUMENTANDTEXTPROCESSING, Deficient mouse, Medicine, lcsh:RC581-607, business, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 030215 immunology
Abstract: retroviruses, retroelements and their hosts Meet ing abstracts - A single PDF containing all abstracts in this Supplement is available here . http://www. biomedcentral.co m/content/pdf/17 42-4690-6-S2-inf o.pdf
Published: 2009

44. Fine characterization of intrahepatic NK cells expressing natural killer receptors in chronic hepatitis B and C

Author: Bonorino, Paula, Ramzan, Muhammad, Camous, Xavier, Dufeu-Duchesne, Tania, Thélu, Marie-Ange, Sturm, Nathalie, Dariz, Aurélie, Guillermet, Christiane, Pernollet, Martine, Zarski, Jean-Pierre, Marche, Patrice, Leroy, Vincent, Jouvin-Marche, Evelyne, 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), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by grants from INSERM , from the Agence Nationale pour la Recherche sur le Sida (ANRS) and from Pole de compétitivité LyonBiopole. P.B. was supported by ANRS, and M.R. from Higher Education Commission of Pakistan and X.C. by Région-Rhône Alpes 'Cluster 10'.
Subjects: MESH: Adolescent, MESH: Killer Cells, Natural, MESH: Middle Aged, MESH: Humans, MESH: Antigens, CD3, MESH: Cell Count, MESH: Hepatitis B, Chronic, MESH: Biological Markers, virus diseases, hemic and immune systems, chemical and pharmacologic phenomena, MESH: Adult, MESH: Antigens, CD56, digestive system diseases, MESH: Male, MESH: Perforin, MESH: Receptors, Natural Killer Cell, MESH: Hepatitis C, Chronic, MESH: NK Cell Lectin-Like Receptor Subfamily C, MESH: Young Adult, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Female, MESH: Liver
Abstract: International audience; BACKGROUND/AIMS: The fate of intrahepatic NK cell subsets in the course of HCV and HBV infections is not clearly understood. METHODS: Blood and intrahepatic CD56(+) NK cell subsets (expressing NKG2A, CD158a,h or CD158b,j receptors) from HCV or HBV patients were quantified by flow cytometry and localized by immunohistochemistry in liver biopsies. RESULTS: A significant reduction in NK cell frequency and a quantitative imbalance between CD56(bright) and CD56(dim) subsets were observed in chronic HCV patients as compared to HBV patients, underlining that the inflammatory environment is not the only cause of these phenomena. The proportions of intrahepatic NK cells expressing either NKG2A, and/or CD158a,h, CD158b,j differed significantly between HCV and HBV patients. A higher frequency of perforin among intrahepatic CD56(+)CD3(-) cells was observed in HCV compared to HBV patients. Double immunohistochemical staining showed that CD56(+)CD3(-) cells were localized within necrotic areas. Immune monitoring of circulating CD56 subsets revealed that CD3(-)CD56(bright)NKG2A(+) and CD3(-)CD56(dim)NKG2A(+) cells were positively correlated with the necroinflammatory score and inversely correlated with viral load, respectively, in HCV patients. CONCLUSIONS: HCV and HBV affect NK cell subsets according to the status of the diseases, especially CD3(-)CD56(dim)NKG2A(+) and CD3(-)CD56(bright)NKG2A(+) cells, may be of interest for disease monitoring.
Published: 2009

45. Individual blood-cell capture and 2D organization on microarrays.: Individual Blood Cell Capture and 2D-Organization on Microarrays

Author: Roupioz, Yoann, Berthet-Duroure, Nathalie, Leichle, Thierry, Pourciel, Jean Bernard, Mailley, Pascal, Cortes, Sandra, Villiers, Marie-Bernadette, Marche, Patrice, Livache, Thierry, Nicu, Liviu, Structures et propriétés d'architectures moléculaire (SPRAM - UMR 5819), Institut Nanosciences et Cryogénie (INAC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'analyse et d'architecture des systèmes (LAAS), Université Toulouse Capitole (UT Capitole), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT), 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), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was partly supported by the French Nanobio Funding program., Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Marche, Patrice, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP)
Subjects: Blood cells, MESH: Humans, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Protein Array Analysis, [SPI.NANO] Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, MESH: Blood Cells, micro-cantilever, surface patterning, MESH: Equipment Failure Analysis, MESH: Flow Cytometry, cell microarray, MESH: Cell Separation, MESH: Sensitivity and Specificity, MESH: Reproducibility of Results, electrochemistry, MESH: Microfluidic Analytical Techniques, MESH: Blood Proteins, [SDV.IMM]Life Sciences [q-bio]/Immunology, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, MESH: Nanotechnology, ComputingMilieux_MISCELLANEOUS, MESH: Cells, Cultured, MESH: Equipment Design
Abstract: International audience
Published: 2009

46. From secretome analysis to immunology: chitosan induces major alterations in the activation of dendritic cells via a TLR4-dependent mechanism

Author: Villiers, Christian, Chevallet, Mireille, Diemer, Hélène, Couderc, Rachel, Freitas, Heidi, Van Dorsselaer, Alain, Marche, Patrice, Rabilloud, Thierry, Rabilloud, Thierry, 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), Institut Pluridisciplinaire Hubert Curien (IPHC), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), and Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Genomics (q-bio.GN), Chitosan, Research, Molecular Sequence Data, Dendritic Cells, Flow Cytometry, Lymphocyte Activation, Mice, Inbred C57BL, Toll-Like Receptor 4, Mice, FOS: Biological sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Animals, Quantitative Biology - Genomics, Amino Acid Sequence, Isoelectric Focusing, Peptides
Abstract: International audience; Dendritic cells are known to be activated by a wide range of microbial products, leading to cytokine production and increased levels of membrane markers such as major histocompatibility complex class II molecules. Such activated dendritic cells possess the capacity to activate naïve T cells. In the present study we demonstrated that immature dendritic cells secrete both the YM1 lectin and lipocalin-2. By testing the ligands of these two proteins, chitosan and siderophores, respectively, we also demonstrated that chitosan, a degradation product of various fungal and protozoal cell walls, induces an activation of dendritic cells at the membrane level, as shown by the up-regulation of membrane proteins such as class II molecules, CD80 and CD86 via a TLR4-dependent mechanism, but is not able to induce cytokine production. This led to the production of activated dendritic cells unable to stimulate T cells. However, costimulation with other microbial products overcame this partial activation and restored the capacity of these activated dendritic cells to stimulate T cells. In addition, successive stimulation with chitosan and then by lipopolysaccharide induced a dose-dependent change in the cytokinic IL-12/IL-10 balance produced by the dendritic cells.
Published: 2009

47. Critical stresses for cancer cell detachment in microchannels

Author: Cécile Couzon, Claude Verdier, Alain Duperray, Verdier, Claude, Laboratoire de Spectrométrie Physique (LSP), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), and Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Materials science, MESH: Cell Line, Tumor, Critical stress, Microfluidics, Biophysics, [SDV.CAN]Life Sciences [q-bio]/Cancer, Models, Biological, MESH: Cell Adhesion, Focal adhesion, [SDV.CAN] Life Sciences [q-bio]/Cancer, MESH: Computer Simulation, Cell Movement, [SPI.MECA.BIOM] Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Cell Line, Tumor, Shear stress, Cell Adhesion, Humans, Computer Simulation, Composite material, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], MESH: Cell Movement, Shearing (physics), Focal Adhesions, MESH: Humans, MESH: Stress, Mechanical, critical stress, [PHYS.MECA.BIOM] Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], MESH: Models, Biological, MESH: Focal Adhesions, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], General Medicine, MESH: Urinary Bladder Neoplasms, Shear (geology), Urinary Bladder Neoplasms, cancer cells, MESH: Shear Strength, Adhesive, Stress, Mechanical, Contact area, Shear Strength
Abstract: International audience; We present experiments involving cancer cells adhering to microchannels, subjected to increasing shear stresses (0.1-30 Pa). Morphological studies were carried out at different shear stresses. Cells exhibit spreading patterns similar to those observed under static conditions, as long as the shear stress is not too high. At critical wall shear stresses (around 2-5 Pa), cell-substrate contact area decreases until detachment at the larger stresses. Critical shear stresses are found to be lower for higher confinements (i.e. smaller cell height to channel height ratio). Fluorescent techniques were used to locate focal adhesions (typically 1 lm(2) in size) under various shearing conditions, showing that cells increase the number of focal contacts in the region facing the flow. To analyze such data, we propose a model to determine the critical stress, resulting from the competition between hydrodynamic forces and the adhesive cell resistance. With this model, typical adhesive stresses exerted at each focal contact can be determined and are in agreement with previous works.
Published: 2009

48. A lymphotoxin-driven pathway to hepatocellular carcinoma.: Sustained hepatic LT signaling causes HCC

Author: Haybaeck, Johannes, Zeller, Nicolas, Wolf, Monika Julia, Weber, Achim, Wagner, Ulrich, Kurrer, Michael Odo, Bremer, Juliane, Lezzi, Giandomenica, Graf, Rolf, Clavien, Pierre-Alain, Thimme, Robert, Blum, Hubert, Nedospasov, Sergei A., Zatloukal, Kurt, Ramzan, Muhammad, Ciesek, Sandra, Pietschmann, Thomas, Marche, Patrice, Karin, Michael, Kopf, Manfred, Browning, Jeffrey, Aguzzi, Adriano, Heikenwalder, Mathias, Marche, Patrice, Institute of Neuropathology, University hospital of Zurich [Zurich], Institutes of Neuropathology and Clinical Pathology, Functional Genomics Center Zurich, Universität Zürich [Zürich] = University of Zurich (UZH)- Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Department of Pathology, Aarau Catonal Hospital, Institute of Integrative Biology, Molecular Biomedicine, Swiss Hepato-Pancreatico-Biliary Center, Department of Internal Medicine, University of Freiburg [Freiburg], Engelhardt Institute of Molecular Biology (ISTC), Russian Academy of Sciences [Moscow] (RAS), Institute of Pathology, Medical University Graz, 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), Division of Experimental Virology, Centre for Experimental and Clinical Infection Research (TWINCORE), Helmholtz Centre for Infection Research (HZI)-Medizinische Hochschule Hannover (MHH)-Helmholtz Centre for Infection Research (HZI)-Medizinische Hochschule Hannover (MHH), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), University of California [San Diego] (UC San Diego), University of California (UC), Department of Immunobiology, Biogen [Cambridge, USA], and Part of the project was achieved with support of the Agence Nationale pour la Recherche sur le Sida (ANRS) and from Pole de Compétitivité LyonBiopole, France. MR was supported by the Higher Education Commission of Pakistan.
Subjects: [SDV.IMM] Life Sciences [q-bio]/Immunology, [SDV.CAN] Life Sciences [q-bio]/Cancer, [SDV.IMM]Life Sciences [q-bio]/Immunology, [SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, neoplasms, digestive system diseases, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology
Abstract: International audience; Hepatitis B and C viruses (HBV and HCV) cause chronic hepatitis and hepatocellular carcinoma (HCC) by poorly understood mechanisms. We show that cytokines lymphotoxin (LT) alpha and beta and their receptor (LTbetaR) are upregulated in HBV- or HCV-induced hepatitis and HCC. Liver-specific LTalphabeta expression in mice induces liver inflammation and HCC, causally linking hepatic LT overexpression to hepatitis and HCC. Development of HCC, composed in part of A6(+) oval cells, depends on lymphocytes and IKappa B kinase beta expressed by hepatocytes but is independent of TNFR1. In vivo LTbetaR stimulation implicates hepatocytes as the major LT-responsive liver cells, and LTbetaR inhibition in LTalphabeta-transgenic mice with hepatitis suppresses HCC formation. Thus, sustained LT signaling represents a pathway involved in hepatitis-induced HCC.
Published: 2009

49. TRACTION PATTERNS OF TUMOR CELLS

Author: Davide Carlo Ambrosi, Claude Verdier, Alain Duperray, Valentina Peschetola, Dipartimento di Matematica 'Giuseppe Peano' [Torino], Università degli studi di Torino (UNITO), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Laboratoire de Spectrométrie Physique (LSP), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), réseau européen RTN, and Politecnico di Torino
Subjects: tumor, Field (physics), Traction (engineering), Finite Element Analysis, Acrylic Resins, Geometry, cell traction, Models, Biological, Quantitative Biology::Cell Behavior, Cell Movement, Neoplasms, Humans, [PHYS.MECA.BIOM]Physics [physics]/Mechanics [physics]/Biomechanics [physics.med-ph], Mathematics, Fourier Analysis, Applied Mathematics, Mathematical analysis, Linear elasticity, inverse problem, elasticity, [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph], Numerical Analysis, Computer-Assisted, Elasticity (physics), Inverse problem, Flow Cytometry, Agricultural and Biological Sciences (miscellaneous), Elliptic partial differential equation, Adjoint equation, Modeling and Simulation, Displacement field
Abstract: International audience; The traction exerted by a cell on a planar deformable substrate can be in- directly obtained on the basis of the displacement field of the underlying layer. The usual methodology used to address this inverse problem is based on the exploitation of the Green tensor of the linear elasticity problem in a half space (Boussinesq problem), coupled with a minimization algorithm under force penalization. A possible alternative strategy is to exploit an adjoint equation, obtained on the basis of a suitable minimiza- tion requirement. The resulting system of coupled elliptic partial differential equations is applied here to determine the force field per unit surface generated by T24 tumor cells on a polyacrylamide substrate. The shear stress obtained by numerical integration provides quantitative insight of the traction field and is a promising tool to investigate the spatial pattern of force per unit surface generated in cell motion, particularly in the case of such cancer cells.
Published: 2009

50. Polypyrrole–Peptide Microarray for Biomolecular Interaction Analysis by SPR Imaging

Author: Andre Roget, Sandra Cortès, Thierry Livache, Patrice N. Marche, Carine Brakha, Marie-Bernadette Villiers, Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), INSERM U823, équipe 8 (Immunologie Analytique des Pathologies Chroniques), Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Structures et propriétés d'architectures moléculaire (SPRAM - UMR 5819), Institut Nanosciences et Cryogénie (INAC), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut de Chimie du CNRS (INC)-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 ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Structures et propriétés d'architectures moléculaire ( SPRAM - UMR 5819 ), Institut Nanosciences et Cryogénie ( INAC ), Université Grenoble Alpes ( UGA ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Grenoble Alpes ( UGA ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Centre National de la Recherche Scientifique ( CNRS ), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Duperray, Alain
Subjects: Microarray, Polymers, Protein Array Analysis, Nanotechnology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Polypyrrole, Models, Biological, Antibodies, Article, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Protein Interaction Mapping, Animals, Humans, Pyrroles, In patient, Surface plasmon resonance, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Immunosorbent Techniques, 030304 developmental biology, 0303 health sciences, Spr imaging, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, Surface Plasmon Resonance, Molecular biology, chemistry, Gold surface, Peptide microarray, Peptides, 030217 neurology & neurosurgery
Abstract: International audience; Nowadays, high-throughput analysis of biological events is a great challenge which could take benefit of the recent development of microarray devices. The great potential of such technology is related to the availability of a chip bearing a large set of probes, stable and easy to obtain, and suitable for ligand-binding detection. Here, we describe a new method based on polypyrrole chemistry, allowing the covalent immobilization of peptides in a microarray format and on a gold surface compatible with the use of surface plasmon resonance. This technique is then illustrated by the detection and characterization of antibodies induced by hepatitis C virus and present in patients' serums.
Published: 2009

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