Search

Your search keyword '"Immunité et cancer ( U932 )"' showing total 325 results
Start Over Author "Immunité et cancer ( U932 )"
325 results on '"Immunité et cancer ( U932 )"'

301. Why the need and how to approach the functional diversity of extracellular vesicles

Author

Tkach, Mercedes, Kowal, Joanna, Théry, Clotilde, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and thery, clotilde

Subjects
0301 basic medicine, Cell signaling, [SDV]Life Sciences [q-bio], Cell Communication, exosomes, Biology, Extracellular vesicles, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Functional diversity, 0302 clinical medicine, Tumor Microenvironment, medicine, Humans, cancer, multi-vesicular endosomes, Tumor microenvironment, Cancer, Articles, medicine.disease, Microvesicles, Cell biology, [SDV] Life Sciences [q-bio], Opinion Piece, 030104 developmental biology, 030220 oncology & carcinogenesis, extracellular vesicles, General Agricultural and Biological Sciences
Abstract

In the past decade, cell-to-cell communication mediated by exosomes has attracted growing attention from biomedical scientists and physicians, leading to several recent publications in top-tier journals. Exosomes are generally defined as secreted membrane vesicles, or extracellular vesicles (EVs), corresponding to the intraluminal vesicles of late endosomal compartments, which are secreted upon fusion of multi-vesicular endosomes with the cell's plasma membrane. Cells, however, were shown to release other types of EVs, for instance, by direct budding off their plasma membrane. Some of these EVs share with exosomes major biophysical and biochemical characteristics, such as size, density and membrane orientation, which impose difficulties in their efficient separation. Despite frequent claims in the literature, whether exosomes really display more important patho/physiological functions, or are endowed with higher potential as diagnostic or therapeutic tools than other EVs, is not yet convincingly demonstrated. In this opinion article, we describe reasons for this lack of precision knowledge in the current stage of the EV field, we review recently described approaches to overcome these caveats, and we propose ways to improve our knowledge on the respective functions of distinct EVs, which will be crucial for future development of well-designed EV-based clinical applications. This article is part of the discussion meeting issue ‘Extracellular vesicles and the tumour microenvironment’.

Published
2017
Full Text
View/download PDF

302. Lymphoid Gene Upregulation on Circulating Progenitors Participates in Their T-Lineage Commitment

Author

Agnès Legrand, Vanessa Zepponi, Valérie Pasqualetto, Amine Boudil, Sophie Ezine, Carolina Martinez-Cingolani, Flora Zavala, Laetitia Gautreau, Jérôme Mégret, Victoria Michaels Lopez, Lamia Skhiri, Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Immunité et cancer (U932), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie-Université Paris Descartes - Paris 5 (UPD5), and Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Male, Receptors, CCR7, T cell, T-Lymphocytes, Immunology, Population, Mice, Nude, Spleen, Bone Marrow Cells, Biology, 03 medical and health sciences, Mice, Receptors, CCR, 0302 clinical medicine, medicine, Immunology and Allergy, Animals, Cell Lineage, IL-2 receptor, Progenitor cell, Receptor, Notch1, Interleukin-7 receptor, education, 030304 developmental biology, Cell Proliferation, 0303 health sciences, education.field_of_study, B-Lymphocytes, Receptors, Interleukin-7, Gene Expression Profiling, Multipotent Stem Cells, GATA3, Interleukin-2 Receptor alpha Subunit, Gene Expression Regulation, Developmental, Cell Differentiation, Molecular biology, Mice, Inbred C57BL, Proto-Oncogene Proteins c-kit, medicine.anatomical_structure, fms-Like Tyrosine Kinase 3, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, Bone marrow, Single-Cell Analysis, 030215 immunology
Abstract

Extrathymic T cell precursors can be detected in many tissues and represent an immediately competent population for rapid T cell reconstitution in the event of immunodeficiencies. Blood T cell progenitors have been detected, but their source in the bone marrow (BM) remains unclear. Prospective purification of BM-resident and circulating progenitors, together with RT-PCR single-cell analysis, was used to evaluate and compare multipotent progenitors (MPPs) and common lymphoid progenitors (CLPs). Molecular analysis of circulating progenitors in comparison with BM-resident progenitors revealed that CCR9+ progenitors are more abundant in the blood than CCR7+ progenitors. Second, although Flt3− CLPs are less common in the BM, they are abundant in the blood and have reduced Cd25+-expressing cells and downregulated c-Kit and IL-7Rα intensities. Third, in contrast, stage 3 MPP (MPP3) cells, the unique circulating MPP subset, have upregulated Il7r, Gata3, and Notch1 in comparison with BM-resident counterparts. Evaluation of the populations’ respective abilities to generate splenic T cell precursors (Lin−Thy1.2+CD25+IL7Rα+) after grafting recipient nude mice revealed that MPP3 cells were the most effective subset (relative to CLPs). Although several lymphoid genes are expressed by MPP3 cells and Flt3− CLPs, the latter only give rise to B cells in the spleen, and Notch1 expression level is not modulated in the blood, as for MPP3 cells. We conclude that CLPs have reached the point where they cannot be a Notch1 target, a limiting condition on the path to T cell engagement.

Published
2014
Full Text
View/download PDF

303. Potential limitations of IL-2 administration for the treatment of experimental acute graft-versus-host disease

Author

Gaëlle H. Martin, Sébastien Maury, Louis Pérol, José L. Cohen, Eliane Piaggio, Immunologie - Immunopathologie - Immunothérapeutique (I3), 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), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Curie [Paris], INSERM U955, équipe 7, Hôpital Henri Mondor, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), APHP Henri Mondor, INSERM Center of Clinical Investigation, Administateur, HAL Sorbonne Université, and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-IFR10-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-IFR10

Subjects
Interleukin 2, Bone marrow transplantation, [SDV.IMM] Life Sciences [q-bio]/Immunology, Regulatory T cell, medicine.medical_treatment, Immunology, Graft vs Host Disease, Antineoplastic Agents, Mice, Transgenic, chemical and pharmacologic phenomena, medicine.disease_cause, Peripheral blood mononuclear cell, T-Lymphocytes, Regulatory, Autoimmunity, 03 medical and health sciences, Mice, 0302 clinical medicine, Immunology and Allergy, Medicine, Animals, Humans, Receptor, 030304 developmental biology, Sirolimus, 0303 health sciences, business.industry, Hematopoietic Stem Cell Transplantation, Interleukin-2 Receptor alpha Subunit, FOXP3, Immunotherapy, Allografts, 3. Good health, Disease Models, Animal, medicine.anatomical_structure, Cytokine, surgical procedures, operative, Graft-vs-host disease, Acute Disease, Heterografts, Interleukin-2, [SDV.IMM]Life Sciences [q-bio]/Immunology, business, Immunosuppressive Agents, 030215 immunology, medicine.drug
Abstract

International audience; Low-dose IL-2 administration can control autoimmunity by specifically activating CD4+ FoxP3+ regulatory T cells (Tregs). Here, we studied IL-2-based immunotherapy in experimental graft-versus-host disease (GVHD). IL-2 administration to donor mice induced a dose-dependent expansion of Tregs in the graft but was insufficient to control GVHD. IL-2 administration to allogeneic-grafted recipient mice activated T-conventional cells (Tcons) and did not prevent GVHD. This loss of IL-2 selectivity toward Tregs was explained by an IL-2-induced increase in the IL-2 receptor α-chain expression on Tcons. Finally, in xeno-GVHD generated by human PBMCs transplanted into immunodeficient mice, low-dose IL-2 increased Treg frequencies but did neither control pro-inflammatory cytokine production by pathogenic Tcons, nor prevented GVHD. Furthermore, combination of low-dose IL-2 with rapamycin was ineffective in this model. Our results indicate that limitations on the use of IL-2 during acute GVHD are likely due to the massive activation of the allogeneic T cells unique to this setting.

Published
2014
Full Text
View/download PDF

304. Biologie-Géologie BCPST-VETO 2e année

Author

Godinot, Claire, Denoeud, Julie, Guipponi, Olivier, Moreau, Hélène, Paulhiac-Pison, Marine, Pons, Marie-Laure, Tejedor, Fanny, Ecole Normale Supérieure Paris-Saclay (ENS Paris Saclay), Centre Scientifique de Monaco (CSM), Institut des sciences cognitives Marc Jeannerod - Centre de neuroscience cognitive - UMR5229 (ISC-MJ), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL), Laboratoire de Minéralogie et Cosmochimie du Muséum (LMCM), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Centre européen de recherche et d'enseignement des géosciences de l'environnement (CEREGE), and Institut de Recherche pour le Développement (IRD)-Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Collège de France (CdF (institution))-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDE]Environmental Sciences
Abstract

International audience

Published
2014

305. Phosphoantigens and butyrophilin 3A1 induce similar intracellular activation signaling in human TCRVγ9+ γδ T lymphocytes

Author

Emilie Decaup, Daniel Olive, Mary Poupot, Caroline Duault, Jean-Jacques Fournié, Christine Bezombes, Ariel Savina, Centre de Recherches en Cancérologie de Toulouse (CRCT), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche en Cancérologie de Marseille (CRCM), Aix Marseille Université (AMU)-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie-Institut National de la Santé et de la Recherche Médicale (INSERM), and Poupot, Mary

Subjects
[SDV.MHEP.HEM] Life Sciences [q-bio]/Human health and pathology/Hematology, medicine.drug_class, medicine.medical_treatment, Immunology, T lymphocytes, Activation, Biology, Signal transduction, Monoclonal antibody, Lymphocyte Activation, Cell Line, Immunophenotyping, 03 medical and health sciences, 0302 clinical medicine, Butyrophilin, Cancer immunotherapy, Antigens, CD, T-Lymphocyte Subsets, medicine, Immunology and Allergy, Humans, Cell barcoding, Protein kinase B, 030304 developmental biology, 0303 health sciences, Mapk p38, Butyrophilins, ZAP70, Antibodies, Monoclonal, Receptors, Antigen, T-Cell, gamma-delta, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, Phosphoproteins, Molecular biology, Cell biology, Antigens, Surface, Phosphoflow cytometry, Gamma-delta, Intracellular, 030215 immunology
Abstract

International audience; Human γδ cells expressing TCRVγ9 are T lymphocytes with great potential for cancer immunotherapy and unconventional pattern of antigen specificity. These HLA-unrestricted lymphocytes are specifically reactive to non-peptide metabolites (phosphoantigens) and to the butyrophilin 3A (BTN3A/CD277) protein. Whether recognition of such highly different structures trigger the same activation signaling pathway remains unclear, however. Here we combined fluorescent cell barcoding and phosphoflow analysis of TCRVγ9(+) T lymphocytes to compare simultaneously the level of several signaling phosphoproteins after activation by phosphoantigen (BrHPP) or by anti-BTN3A (monoclonal antibody 20.1). This approach shows that the same pathways involving ZAP70, PLCγ2, Akt, NFκB p65, MAPK p38 and Erk1, were induced by either of these stimuli. These data strongly suggest the TCRVγ9(+) T lymphocytes detect phosphoantigens and butyrophilin A3 by the same recognition process.

Published
2014
Full Text
View/download PDF

306. Biogenesis and secretion of exosomes

Author

Joanna Kowal, Mercedes Tkach, Clotilde Théry, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Exosome-related work performed in the Théry lab is supported by fundings from Institut National de la Santé et de la RechercheMédicale, Institut Curie (including fellowship to J. Kowal), Association pour laRecherche contre le Cancer (including fellowship to M. Tkach), Fondation de France, Agence Nationale de la Recherche ([ANR-10-IDEX-0001-02 PSL*] and [ANR-11-LABX-0043])., ANR-10-IDEX-0001,PSL,Paris Sciences et Lettres(2010), ANR-10-IDEX-0003,IDEX BORDEAUX,Initiative d'excellence de l'Université de Bordeaux(2010), thery, clotilde, Initiative d'excellence - Paris Sciences et Lettres - - PSL2010 - ANR-10-IDEX-0001 - IDEX - VALID, and Initiative d'excellence de l'Université de Bordeaux - - IDEX BORDEAUX2010 - ANR-10-IDEX-0003 - IDEX - VALID

Subjects
Vesicle, [SDV]Life Sciences [q-bio], Cell Membrane, Biological Transport, Cell Biology, Biology, Exosomes, Extracellular vesicles, Microvesicles, Cell biology, [SDV] Life Sciences [q-bio], Animals, Humans, Secretion, Intracellular, Biogenesis, Scientific society
Abstract

International audience; Although observed for several decades, the release of membrane-enclosed vesicles by cells into their surrounding environment has been the subject of increasing interest in the past few years, which led to the creation, in 2012, of a scientific society dedicated to the subject: the International Society for Extracellular Vesicles. Convincing evidence that vesicles allow exchange of complex information fuelled this rise in interest. But it has also become clear that different types of secreted vesicles co-exist, with different intracellular origins and modes of formation, and thus probably different compositions and functions. Exosomes are one sub-type of secreted vesicles. They form inside eukaryotic cells in multivesicular compartments, and are secreted when these compartments fuse with the plasma membrane. Interestingly, different families of molecules have been shown to allow intracellular formation of exosomes and their subsequent secretion, which suggests that even among exosomes different sub-types exist.

Published
2014
Full Text
View/download PDF

307. Le VIH-2 révèle un mécanisme antiviral de détection par l’immunité innée

Author

Nicolas Manel, Xavier Lahaye, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and LAHAYE, Xavier

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], General Medicine, Biology, Molecular biology, General Biochemistry, Genetics and Molecular Biology, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2014
Full Text
View/download PDF

308. Biogenesis, Secretion, and Intercellular Interactions of Exosomes and Other Extracellular Vesicles

Author

Graça Raposo, Marina Colombo, Clotilde Théry, thery, clotilde, Compartimentation et dynamique cellulaires (CDC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Curie [Paris]-Centre National de la Recherche Scientifique (CNRS), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Université Pierre et Marie Curie - Paris 6 (UPMC)

Subjects
Cell type, Reticulocytes, Endosome, [SDV]Life Sciences [q-bio], Cytological Techniques, Cell Communication, Endosomes, Biology, Exosomes, Membrane Fusion, Membrane Lipids, Cell-Derived Microparticles, Neoplasms, Centrifugation, Density Gradient, Animals, Humans, Secretion, RNA, Messenger, Transport Vesicles, ComputingMilieux_MISCELLANEOUS, B-Lymphocytes, Vesicle, Membrane Proteins, Biological Transport, Extracellular Fluid, Cell Biology, Extracellular vesicle, Microvesicles, Cell biology, [SDV] Life Sciences [q-bio], MicroRNAs, Eukaryotic Cells, Prokaryotic Cells, rab GTP-Binding Proteins, SNARE Proteins, Biogenesis, Intracellular, Developmental Biology
Abstract

In the 1980s, exosomes were described as vesicles of endosomal origin secreted from reticulocytes. Interest increased around these extracellular vesicles, as they appeared to participate in several cellular processes. Exosomes bear proteins, lipids, and RNAs, mediating intercellular communication between different cell types in the body, and thus affecting normal and pathological conditions. Only recently, scientists acknowledged the difficulty of separating exosomes from other types of extracellular vesicles, which precludes a clear attribution of a particular function to the different types of secreted vesicles. To shed light into this complex but expanding field of science, this review focuses on the definition of exosomes and other secreted extracellular vesicles. Their biogenesis, their secretion, and their subsequent fate are discussed, as their functions rely on these important processes.

Published
2014
Full Text
View/download PDF

309. Unexpected remote effect in red fluorescent sensors based on extended APTRA

Author

Mayeul Collot, Rodolphe Vuilleumier, Aurélie Lasoroski, Alsu I. Zamaleeva, Jean-Maurice Mallet, Anne Feltz, Université Pierre et Marie Curie - Paris 6 (UPMC), Processus d'Activation Sélective par Transfert d'Energie Uni-électronique ou Radiatif (UMR 8640) (PASTEUR), Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie de Coordination des Eléments f (LCF) (LCCEf), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie de l'ENS Paris (UMR 8197/1024) (IBENS), Département de Biologie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Agence Nationale de la Recherche (ANR), Institut de biologie de l'ENS Paris (IBENS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Biologie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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), École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS Paris)-École normale supérieure - Paris (ENS Paris)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie-Université Paris Descartes - Paris 5 (UPD5)

Subjects
Coordination sphere, 010405 organic chemistry, Chemistry, Stereochemistry, [SDV]Life Sciences [q-bio], Organic Chemistry, Metal sensor, 010402 general chemistry, 01 natural sciences, Biochemistry, Fluorescence, DFT, 0104 chemical sciences, X-Rhodamine, Crystallography, Aminophenol triacetic acid, Fluorescent probe, Remote control, Drug Discovery, Click chemistry, Side chain, Moiety, [CHIM]Chemical Sciences, Chelation, Order of magnitude
Abstract

International audience; Herein is described the synthesis and spectroscopic characterizations of three new OFF ON red-emitting and water-soluble sensors, CAXR (Clicked APTRA X-Rhodamine). These dyes are based on an extended APTRA (aminophenol triacetic acid) motif. Three different side chains were added by click chemistry in order to complete the coordination sphere with a chelate moiety composed of a triazolyl and an iminol. The fluorescent response (F/F-0) of these probes follows the order: Cd2+>Zn2+>Pb2+>Hg2+. An important and unexpected effect of the side chain structure on the Kd was observed (up to one order of magnitude, Cadmium Kd from 252 to 21 mu M). This remote effect of the side chains was studied by DFT calculations and was attributed to a twisted conformation of the CAXR-Py:Cd2+ complex.

Published
2013
Full Text
View/download PDF

310. Effets différentiels de la cytokine lymphopoietine stromale thymique sur les sous-populations de cellules dendritiques humaines

Author

Martinez Cingolani, Carolina, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Sud - Paris XI, Vassili Soumelis, and STAR, ABES

Subjects
Chimiokines, Subsets, Lymphopoietine stromale thymique, Thymic stromal lymphopoietin, [SDV.IMM] Life Sciences [q-bio]/Immunology, Cellules dendritiques, Humain, [SDV.IMM]Life Sciences [q-bio]/Immunology, Chemokines, Dendritic cells, Sous-populations, Migration, Human
Abstract

Once activated, Dendritic Cells (DCs) migrate to the lymphoid organs and exert their role as professional antigen presenting cells. They are able to induce the activation and differentiation of naïve T cells into different types of T helper cells. The T cell response must be suited to the type of inflammation. This is ensured by DCs at two levels. First DCs are functionally plastic. This means that their behavior is subdued to the integrated signals coming from the inflammatory microenvironment. Secondly, the DC population is diverse. Indeed, different DC subsets have different functional specializations. My thesis was focused on the differential response of human DC subsets to Thymic stromal lymphopoietin (TSLP). This cytokine is secreted by inflamed skin and epithelia, and strongly activates myeloid DCs. The TSLP-activated DCs secrete the inflammatory chemokines CCL17 and CCL22, prime an inflammatory Th2 response, and have been involved in the pathogenesis of allergic inflammation. By systematically comparing the response of human blood BDCA-1+ and BDCA-3+ DCs to TSLP stimulation we found that both of these DC subsets get activated by TSLP. However TSLP synergizes with TGF-β to induce the differentiation of blood BDCA-1+ and not BDCA-3+ DCs into Langerhans Cells. Moreover, TSLP induces cell migration and chemokine secretion only on the blood BDCA-1+ subset. Further analysis of the mechanisms implicated in TSLP-induced DC migration revealed that TSLP is required to induce DC migration, but this effect is dependent on the expression of a PTX-sensitive chemokine receptor. Overall our results reveal new precursor capacities of blood DC subsets, different functional properties of blood DC subsets stimulated by TSLP and highlight intricate mechanisms underlying TSLP-induced DC migration., Une fois activées, les cellules dendritiques (DCs) migrent dans les organes lymphoïdes ou elles exercent leur rôle de cellules présentatrices d’antigène professionnelles. Elles sont capables d’activer et d’induire la différenciation des lymphocytes T naïfs en différentes sous-populations de lymphocytes T auxiliaires. L’ajustement de la réponse lymphocytaire T au type d’inflammation est assuré par les DCs à deux niveaux. Premièrement, grâce à leur plasticité, les DCs adaptent leur comportement en fonction de la combinaison de signaux issus du microenvironnement inflammatoire. Deuxièmement, il existe différentes sous-populations de cellules dendritiques ayant de différentes spécialisations fonctionnelles.Mon travail de thèse s’est concentré sur l’étude de la diversité des réponses des sous-populations de cellules dendritiques humaines suite à la stimulation par la cytokine lymphopoïetine stromale thymique (TSLP). Cette cytokine est secrétée par les cellules épithéliales et la peau au cours de l’inflammation. La TSLP active principalement les DCs myéloïdes, induisant celles-ci à secréter les chimiokines inflammatoires CCL17 et CCL22. Les DCs activées par la TSLP (TSLP-DCs) induisent une réponse inflammatoire de type Th2, et sont impliquées dans le développement de l’allergie. La comparaison systématique de la réponse à la TSLP par les sous-populations de DCs du sang, BDCA1+ et BDCA3+, nous a permis de montrer que ces deux sous-populations sont activées par la TSLP. Toutefois, nos résultats montrent que la TSLP, en synergie avec le TGF-β, induit la différenciation des DCs BDCA-1+ en cellules de Langerhans, mais pas celle des DCs BDCA-3+. De plus, la TSLP induit la migration cellulaire et la sécrétion de chimiokines seulement chez la sous-population de DCs BDCA-1+. Des analyses complémentaires des mécanismes impliqués dans la migration des DCs BDCA-1+ en réponse à la TSLP révèlent que, d’une part la TSLP est indispensable à l’induction de la migration et d’autre part, qu’un récepteur de chimiokines, sensible à la Toxine Pertussique serait impliqué. Au final, nos résultats révèlent (i) de nouvelles capacités des DCs en tant que cellules précurseurs, (ii) de différentes propriétés fonctionnelles des sous-populations de DCs en réponse à la stimulation par la TSLP, (iii) et démontrent la complexité des mécanismes impliqués dans la migration des DCs induite par la TSLP.

Published
2013

311. CD36-specific antibodies block release of HIV-1 from infected primary macrophages and its transmission to T cells

Author

Philippe Benaroch, Raphael Gaudin, François-Xavier Gobert, Marc Rabaza-Gairi, Stefano Berre, Bruna Cunha de Alencar, Marion Desdouits, Mélanie Chabaud, Nadia Naffakh, Mabel Jouve, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Institut Pasteur [Paris]-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Work was supported by grants from Agence Nationale de Recherche contre le SIDA (ANRS) and Ensemble contre le SIDA (Sidaction) to P. Benaroch and fellowships to R. Gaudin (ANRS), S. Berre (Curie Institute), M. Desdouits (ANRS), and B. Cunha de Alencar (Fondation pour la Recherche Médicale)., Institut Pasteur [Paris] (IP)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie-Université Paris Descartes - Paris 5 (UPD5), and Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Institut Pasteur [Paris]

Subjects
CD36 Antigens, medicine.drug_class, CD36, T-Lymphocytes, viruses, [SDV]Life Sciences [q-bio], Immunology, Molecular Sequence Data, HIV Core Protein p24, HIV Infections, Biology, Monoclonal antibody, Virus, Article, 03 medical and health sciences, 0302 clinical medicine, Antibody Specificity, Antibodies, Bispecific, parasitic diseases, medicine, Immunology and Allergy, Gene silencing, Humans, Amino Acid Sequence, Scavenger receptor, Antibodies, Blocking, Cells, Cultured, Virus Release, 030304 developmental biology, 0303 health sciences, Macrophages, Virus Assembly, Virion, Antibodies, Monoclonal, Virology, 3. Good health, Lipoproteins, LDL, 030220 oncology & carcinogenesis, biology.protein, HIV-1, Ectopic expression, Antibody, HeLa Cells
Abstract

Antibody targeting CD36 on HIV-1–infected macrophages results in rapid retention of virions within virus-containing compartments to inhibit release and viral transmission to T cells., HIV-1–infected macrophages likely represent viral reservoirs, as they accumulate newly formed virions in internal virus-containing compartments (VCCs). However, the nature and biogenesis of VCCs remain poorly defined. We show that upon HIV-1 infection of primary human macrophages, Gag is recruited to preexisting compartments containing the scavenger receptor CD36, which then become VCCs. Silencing of CD36 in HIV-1–infected macrophages decreases the amount of virions released. Strikingly, soluble anti-CD36 antibodies, but not the natural ligands of CD36, inhibit release of virions from HIV-1–infected macrophages and the transmission of virus to CD4+ T cells. The effect of the antibodies is potent, rapid, and induces the retention of virions within VCCs. Ectopic expression of CD36 in HeLa cells renders them susceptible to the inhibitory effect of the anti-CD36 mAb upon HIV-1 infection. We show that the anti-CD36 mAb inhibits HIV-1 release by clustering newly formed virions at their site of budding, and that signaling via CD36 is not required. Thus, HIV-1 reservoirs in macrophages may be tackled therapeutically using anti-CD36 antibodies to prevent viral dissemination.

Published
2013
Full Text
View/download PDF

312. An Improved Strategy for Easy Process Monitoring and Advanced Purification of Recombinant Proteins

Author

Miladi, Baligh, Dridi, Cyrine, El Marjou, Ahmed, Boeuf, Guilhem, Bouallagui, Hassib, Dufour, Florence, Di Martino, Patrick, Elm’selmi, Abdellatif, Equipe de recherche sur les relations matrice extracellulaire-cellules (ERRMECe), Fédération INSTITUT DES MATÉRIAUX DE CERGY-PONTOISE (I-MAT), Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), Institut National des Sciences Appliquées et de Technologie [Tunis] (INSAT), DI MARTINO, PATRICK, and Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie-Université Paris Descartes - Paris 5 (UPD5)

Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, ComputingMilieux_MISCELLANEOUS, [SDV.BIO] Life Sciences [q-bio]/Biotechnology
Abstract

International audience

Published
2013

313. Utilisation d'un anticorps monoclonal anti-Tn en immunothérapie des cancers

Author

Heitzmann-Daverton, Adèle, STAR, ABES, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université René Descartes - Paris V, and Sebastian Amigorena

Subjects
Monoclonal antibody, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Immunoconjugué, Auristatine F, Immunothérapie, Immunotherapy, Anticorps monoclonal, Internalisation, Saporine, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, Internalization
Abstract

Pas de résumé en anglais, La transformation des cellules normales de l’organisme en un phénotype malin est souvent accompagnée de changements dans leur antigénicité. L’antigène Tn (GalNac-O-Ser/Thréo) est un antigène (Ag) glycopeptidique spécifique des tumeurs et exprimé à la membrane plasmique des cellules cancéreuses dans la majorité des carcinomes humains ainsi que dans certaines tumeurs hématologiques, tandis qu’il n’est pas détecté dans les cellules normales. Il représente donc une cible potentielle très intéressante pour l’immunothérapie passive par anticorps, car il n’est pas détectable dans les cellules normales, mais est démasqué dans environ 90% des cancers épithéliaux du fait d’une dérégulation des processus de glycosylation. Les anticorps monoclonaux (AcM) spécifiques d’antigènes exprimés à la membrane des cellules tumorales ont une efficacité prouvée dans le traitement de certains cancers. Ces AcM thérapeutiques sont particulièrement intéressants pour le traitement des cancers du fait de leur forte spécificité pour les cellules tumorales et de leur faible toxicité pour les cellules normales, contrairement aux chimiothérapies conventionnelles, mais leur mécanisme d’action est encore mal connu. L’AcM Chi-Tn est un anticorps chimérique homme/souris capable de se fixer de façon spécifique à l’antigène tumoral Tn, alors qu’il ne se fixe pas sur les cellules normales. Cet AcM pourrait donc être envisagé comme agent thérapeutique dans le traitement des cancers épithéliaux par immunothérapie passive.Nous nous sommes intéressés à l’AcM Chi-Tn non couplé en vue d’analyser son mécanisme d’action et d’évaluer son efficacité thérapeutique in vivo. Nous avons montré que l’AcM Chi-Tn seul ne possède pas d’effet toxique direct sur les lignées de cellules tumorales Tn-positives in vitro. Cependant, en présence de macrophages, cet AcM est capable d’induire la lyse de ces cellules par un mécanisme d’ADCC. In vivo, l’AcM Chi-Tn, associé à la cyclophosphamide, induit le rejet d'une tumeur du sein dans plus de 80% des souris. Cette inhibition de la croissance tumorale est abolie chez les souris déficientes pour la chaine associée aux récepteurs RFc activateurs, suggérant in vivo un mécanisme d’ADCC. Par l’étude microscopique du microenvironnement tumoral, nous avons observé que les cellules tumorales forment in vivo des synapses avec des macrophages, des neutrophiles, mais aussi des lymphocytes B. Des expériences de survie in vivo chez des souris déficientes pour différentes populations cellulaires montrent que les lymphocytes T semblent nécessaires à la protection des souris par Chi-Tn contre la tumeur. Ainsi, ces résultats confirment le rôle des effecteurs exprimant des RFc activateurs, mais aussi le rôle indispensable de la réponse immune adaptative pour assurer l'effet thérapeutique des AcM.Nous nous sommes également intéressés à l’utilisation potentielle de l’AcM Chi-Tn comme vecteur d’agents cytotoxiques. In vivo, dans un modèle de tumeurs solides chez la souris, des expériences de biodistribution montrent que l’AcM Chi-Tn est capable de cibler spécifiquement les zones tumorales, ce qui en fait un anticorps potentiellement utilisable comme vecteur de molécules toxiques. L’internalisation du complexe anticorps/antigène cible est un pré-requis nécessaire à l’utilisation de l’anticorps conjugué. Nous avons montré in vitro que l’AcM Chi-Tn est internalisé dans les endosomes précoces et de recyclage pendant un temps relativement long, faisant de cet AcM un bon candidat pour être couplé à des agents cytotoxiques. Durant ma thèse, nous avons couplé l’AcM Chi-Tn à la toxine saporine ou à la molécule cytotoxique auristatine F, et nous avons montré in vitro que ces conjugués sont cytotoxiques sur des lignées cellulaires Tn-positives.

Published
2013

314. Rôle des protéines Rab27 dans la sécrétion des exosomes : implication dans l’étude des réponses immunitaires tumorales

Author

Bobrie, Angélique, STAR, ABES, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université René Descartes - Paris V, and Clotilde Théry

Subjects
[SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Protéines Rab27, Immunology, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

Pas de résumé en anglais, Pas de résumé en français

Published
2012

315. WITHDRAWN: Reprint of: A new tagged-TEV protease: Construction, optimisation of production, purification and test activity

Author

Guilhem Boeuf, Ahmed El Marjou, Cyrine Dridi, Patrick Di Martino, Hassib Bouallagui, Abdellatif Elm’selmi, Baligh Miladi, Florence Dufour, Equipe de recherche sur les relations matrice extracellulaire-cellules (ERRMECe), Fédération INSTITUT DES MATÉRIAUX DE CERGY-PONTOISE (I-MAT), Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine, Institut National des Sciences Appliquées et de Technologie [Tunis] (INSAT), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie-Université Paris Descartes - Paris 5 (UPD5), and DI MARTINO, PATRICK

Subjects
0106 biological sciences, Proteases, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, medicine.medical_treatment, medicine.disease_cause, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, 01 natural sciences, law.invention, 03 medical and health sciences, Affinity chromatography, law, 010608 biotechnology, medicine, TEV protease, Escherichia coli, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Protease, biology, Chemistry, Tobacco etch virus, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, biology.organism_classification, Fusion protein, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Biochemistry, Chemical engineering, Recombinant DNA, Biotechnology
Abstract

The Tobacco Etch Virus (TEV) protease is frequently used in the cleavage of recombinant fusion proteins because of its efficiency and high specificity. In this work, we present a new recombinant form of TEV termed Streptag II-TEV for high-level production and purification of TEV protease from Escherichia coli and compare it to the hexahistidine (6xHis) tagged version of TEV. The effects of varying the host strain, the bacterial induction temperature (25, 30 and 37°C) and the IPTG inducer concentration on production and solubility of the two recombinant TEV proteases have been examined. Optimal Streptag II-TEV protein expression were obtained in the E. coli KRX strain under an induction temperature of 25°C in the presence of IPTG at 0.5mM. In these conditions, soluble Streptag II-TEV and 6xHis-TEV proteases accounted for about 25% and 18% of total soluble proteins, respectively. About 70% of Streptag II-TEV and 60% of 6xHis-TEV were detected in the supernatant. Streptag II-TEV protease purifies to near homogeneity (approximately 99%) via a simple, single step Strep-Tactin chromatography purification protocol based on the presence of Streptag II. The higher production of Streptag II-TEV coupled to its purification and cleavage efficiencies make it an attractive alternate to 6xHis-TEV.

Published
2011

316. A new tagged-TEV protease: construction, optimisation of production, purification and test activity

Author

Florence Dufour, Baligh Miladi, Patrick Di Martino, Cyrine Dridi, Ahmed El Marjou, Abdellatif Elm’selmi, Hassib Bouallagui, Guilhem Boeuf, Equipe de recherche sur les relations matrice extracellulaire-cellules (ERRMECe), Fédération INSTITUT DES MATÉRIAUX DE CERGY-PONTOISE (I-MAT), Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine, Institut National des Sciences Appliquées et de Technologie [Tunis] (INSAT), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), DI MARTINO, PATRICK, and Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie-Université Paris Descartes - Paris 5 (UPD5)

Subjects
0106 biological sciences, Proteases, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, medicine.medical_treatment, Recombinant Fusion Proteins, Genetic Vectors, Molecular Sequence Data, Gene Expression, medicine.disease_cause, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, 01 natural sciences, law.invention, Plant Viruses, 03 medical and health sciences, Viral Proteins, Affinity chromatography, law, 010608 biotechnology, Endopeptidases, Tobacco, medicine, TEV protease, Escherichia coli, Histidine, Cloning, Molecular, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Chromatography, Protease, biology, Base Sequence, Chemistry, Tobacco etch virus, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, biology.organism_classification, Fusion protein, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Biochemistry, Recombinant DNA, Oligopeptides, Biotechnology, Plasmids
Abstract

The Tobacco Etch Virus (TEV) protease is frequently used in the cleavage of recombinant fusion proteins because of its efficiency and high specificity. In this work, we present a new recombinant form of TEV termed Streptag II-TEV for high-level production and purification of TEV protease from Escherichia coli and compare it to the hexahistidine (6xHis) tagged version of TEV. The effects of varying the host strain, the bacterial induction temperature (25, 30 and 37°C) and the IPTG inducer concentration on production and solubility of the two recombinant TEV proteases have been examined. Optimal Streptag II-TEV protein expression were obtained in the E. coli KRX strain under an induction temperature of 25°C in the presence of IPTG at 0.5 mM. In these conditions, soluble Streptag II-TEV and 6xHis-TEV proteases accounted for about 25% and 18% of total soluble proteins, respectively. About 70% of Streptag II-TEV and 60% of 6xHis-TEV were detected in the supernatant. Streptag II-TEV protease purifies to near homogeneity (approximately 99%) via a simple, single step Strep-Tactin chromatography purification protocol based on the presence of Streptag II. The higher production of Streptag II-TEV coupled to its purification and cleavage efficiencies make it an attractive alternate to 6xHis-TEV.

Published
2010

317. Hypoxanthine-guanine phosphoribosyl transferase regulates early developmental programming of dopamine neurons: implications for Lesch-Nyhan disease pathogenesis

Author

Alvarez-Fischer, Daniel, Castagner, François, Stettler, Olivier, Massiani-Beaudoin, Olivia, Moya, Kenneth, Bouillot, Colette, Oertel, Wolfgang, Lombès, Anne, Faigle, Wolfgang, Joshi, Rajiv L, Hartmann, Andreas, Prochiantz, Alain, Ceballos-Picot, I., Mockel, L., Potier, M.-C., Dauphinot, L., Shirley, L., Torero-Ibad, R., Fuchs, Julia, Jinnah, H.A., Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Centre National de la Recherche Scientifique (CNRS), Centre interdisciplinaire de recherche en biologie (CIRB), Labex MemoLife, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de biologie physico-chimique (IBPC (FR_550)), Centre National de la Recherche Scientifique (CNRS), Somnomar, Sleep Research Institute, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Mathématiques de Bordeaux (IMB), Université Bordeaux Segalen - Bordeaux 2-Université Sciences et Technologies - Bordeaux 1-Université de Bordeaux (UB)-Institut Polytechnique de Bordeaux (Bordeaux INP)-Centre National de la Recherche Scientifique (CNRS), Chaire Processus morphogénétiques, Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Labex MemoLife, UMR7637, 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)-CHU Pitié-Salpêtrière [APHP], Collège de France (CDF), Collège de France (CdF), Université Paris Descartes - Paris 5 (UPD5)-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)-Institut Curie-Université Paris Descartes - Paris 5 (UPD5), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Collège de France (CdF)-PSL Research University (PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Université Bordeaux Segalen - Bordeaux 2-Université Sciences et Technologies - Bordeaux 1 (UB)-Université de Bordeaux (UB)-Institut Polytechnique de Bordeaux (Bordeaux INP)-Centre National de la Recherche Scientifique (CNRS), Collège de France - Chaire Processus morphogénétiques, and Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Labex MemoLife

Subjects
Male, Hypoxanthine Phosphoribosyltransferase, Lesch-Nyhan Syndrome, cells, Dopamine, Neurogenesis, [SDV]Life Sciences [q-bio], genetic processes, Nerve Tissue Proteins, Biology, Cell Line, 03 medical and health sciences, Mice, 0302 clinical medicine, Genetics, medicine, Animals, Humans, Molecular Biology, Genetics (clinical), Cells, Cultured, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, Regulation of gene expression, Homeodomain Proteins, Neurons, 0303 health sciences, Dopaminergic, nutritional and metabolic diseases, Gene Expression Regulation, Developmental, General Medicine, Articles, Fibroblasts, medicine.disease, Phenotype, engrailed, Cell biology, enzymes and coenzymes (carbohydrates), medicine.anatomical_structure, Hypoxanthine-guanine phosphoribosyltransferase, Dopaminergic pathways, Lesch–Nyhan syndrome, 030217 neurology & neurosurgery
Abstract

Hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency results in Lesch-Nyhan disease (LND), where affected individuals exhibit a characteristic neurobehavioral disorder that has been linked with dysfunction of dopaminergic pathways of the basal ganglia. Since the functions of HPRT, a housekeeping enzyme responsible for recycling purines, have no direct relationships with the dopaminergic pathways, the mechanisms whereby HPRT deficiency affect them remain unknown. The current studies demonstrate that HPRT deficiency influences early developmental processes controlling the dopaminergic phenotype, using several different cell models for HPRT deficiency. Microarray methods and quantitative PCR were applied to 10 different HPRT-deficient (HPRT(-)) sublines derived from the MN9D cell line. Despite the variation inherent in such mutant sublines, several consistent abnormalities were evident. Most notable were increases in the mRNAs for engrailed 1 and 2, transcription factors known to play a key role in the specification and survival of dopamine neurons. The increases in mRNAs were accompanied by increases in engrailed proteins, and restoration of HPRT reverted engrailed expression towards normal levels, demonstrating a functional relationship between HPRT and engrailed. The functional relevance of the abnormal developmental molecular signature of the HPRT(-) MN9D cells was evident in impoverished neurite outgrowth when the cells were forced to differentiate chemically. To verify that these abnormalities were not idiosyncratic to the MN9D line, HPRT(-) sublines from the SK-N-BE(2) M17 human neuroblastoma line were evaluated and an increased expression of engrailed mRNAs was also seen. Over-expression of engrailed occurred even in primary fibroblasts from patients with LND in a manner that suggested a correlation with disease severity. These results provide novel evidence that HPRT deficiency may affect dopaminergic neurons by influencing early developmental mechanisms.

Published
2009
Full Text
View/download PDF

318. OA011-03. Clusterin, a natural ligand of DC-SIGN present in human semen inhibits HIV capture and transmission by dendritic cells

Author

Wolfgang Faigle, Federico Remes Lenicov, Ana Ceballos, Hugues Lortat-Jacob, Franck Fieschi, Fernando Arenzana-Seisdedos, Jean-Claude Michalski, Juan Sabatté, Jorge Geffner, Michel Thépaut, Sebastian Amigorena, Claudia Rodríguez, Willy Morelle, National Reference center for AIDS, Institut de biologie structurale (IBS - UMR 5075), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS), Institut Curie, Immunité et cancer (U932), Institut Curie-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Immunologie Virale, Institut Pasteur [Paris], Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut Curie [Paris], Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Glycobiologie Structurale et Fonctionnelle UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), 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 National de la Recherche Agronomique (INRA)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale ( IBS - UMR 5075 ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Grenoble Alpes ( UGA ) -Université Joseph Fourier - Grenoble 1 ( UJF ), Immunité et cancer ( U932 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Institut Curie, Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 ( UGSF ), Institut National de la Recherche Agronomique ( INRA ) -Université de Lille-Centre National de la Recherche Scientifique ( CNRS ), and BMC, Ed.

Subjects
lcsh:Immunologic diseases. Allergy, 0303 health sciences, Philosophy, Human immunodeficiency virus (HIV), medicine.disease_cause, 3. Good health, 03 medical and health sciences, [ SDV.MHEP.MI ] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, 0302 clinical medicine, Infectious Diseases, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Virology, Immunology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, medicine, Oral Presentation, 030212 general & internal medicine, lcsh:RC581-607, Humanities, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology
Abstract

Address: 1National Reference center for AIDS, Buenos Aires, Argentina, 2Institut Curie, Paris, France, 3Institut de Biologie Structurale, UMR CNRSCEA-UJF, Grenoble, France, 4Institut de Biologie Structurale, UMR CNRS-CEA-UJF, Grenoble, France, 5UMR CNRS 8576, Unite de Glycobiologie Structurale et fonctionnelle, Lille, France, 6Unite d'Immunologie Virale, Institut Pasteur, Paris, France and 7U932 INSERM, Institut Curie, Immunite et Cancer, Paris, France * Corresponding author

Full Text
View/download PDF

319. CD8+ Tumor-Infiltrating T Cells Are Trapped in the Tumor-Dendritic Cell Network

Author

Luc Fetler, Clotilde Théry, Christophe Combadière, Sophie Krumeich, Alexandre Boissonnas, Sebastien Jacquelin, Fabrice Licata, Lucie Poupel, Sebastian Amigorena, CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Immunité et Infection, Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR113-Institut National de la Santé et de la Recherche Médicale (INSERM), Physico-Chimie-Curie (PCC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Curie [Paris]-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d'Immunologie [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), HAL UPMC, Gestionnaire, Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR113-Université Pierre et Marie Curie - Paris 6 (UPMC), Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC), Service d'immunologie [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP], Université Pierre et Marie Curie - Paris 6 ( UPMC ) -IFR113-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Physico-Chimie-Curie ( PCC ), Centre National de la Recherche Scientifique ( CNRS ) -INSTITUT CURIE-Université Pierre et Marie Curie - Paris 6 ( UPMC ), Immunité et cancer ( U932 ), and Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Institut Curie

Subjects
Cancer Research, Pathology, medicine.medical_specialty, Receptors, Antigen, T-Cell, Mice, Transgenic, CD8-Positive T-Lymphocytes, Biology, lcsh:RC254-282, Mice, 03 medical and health sciences, Interleukin 21, Lymphocytes, Tumor-Infiltrating, 0302 clinical medicine, Immune system, [ SDV.MHEP ] Life Sciences [q-bio]/Human health and pathology, Immune Tolerance, medicine, Animals, Cytotoxic T cell, IL-2 receptor, Antigens, Antigen-presenting cell, 030304 developmental biology, 0303 health sciences, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Dendritic Cells, Dendritic cell, Natural killer T cell, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, 3. Good health, Mice, Inbred C57BL, 030220 oncology & carcinogenesis, Female, CD8, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, T-Lymphocytes, Cytotoxic, Research Article
Abstract

International audience; Chemotherapy enhances the antitumor adaptive immune T cell response, but the immunosuppressive tumor environment often dominates, resulting in cancer relapse. Antigen-presenting cells such as tumor-associated macrophages (TAMs) and tumor dendritic cells (TuDCs) are the main protagonists of tumor-infiltrating lymphocyte (TIL) immuno-suppression. TAMs have been widely investigated and are associated with poor prognosis, but the immuno-suppressive activity of TuDCs is less well understood. We performed two-photon imaging of the tumor tissue to examine the spatiotemporal interactions between TILs and TuDCs after chemotherapy. In a strongly immuno-suppressive murine tumor model, cyclophosphamide-mediated chemotherapy transiently enhanced the antitumor activity of adoptively transferred ovalbumin-specific CD8 + T cell receptor transgenic T cells (OTI) but barely affected TuDC compartment within the tumor. Time lapse imaging of living tumor tissue showed that TuDCs are organized as a mesh with dynamic interconnections. Once infiltrated into the tumor parenchyma, OTI T cells make antigen-specific and long-lasting contacts with TuDCs. Extensive analysis of TIL infiltration on histologic section revealed that after chemotherapy the majority of OTI T cells interact with TuDCs and that infiltration is restricted to TuDC-rich areas. We propose that the TuDC network exerts antigen-dependent unproductive retention that trap T cells and limit their antitumor effectiveness.

Full Text
View/download PDF

320. The p53-inducible TSAP6 gene product regulates apoptosis and the cell cycle and interacts with Nix and the Myt1 kinase

Author

Robert Amson, Véronique Bouvard, Marcel Tuynder, Stephanie Morchoisne, Christophe Cans, Aude Roborel de Climens, Sylvie Prieur, Vanessa Nancy-Portebois, Laurent Susini, Brent Passer, Nathalie Amzallag, Giusy Fiucci, Virginie Crible, Adam Telerman, Jean Dausset, Alexandra Lespagnol, Moshe Oren, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie Moléculaire de la Cellule (LBMC), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Organisation Reconstruction Travail - ORT (FRANCE), Centre internationale de recherche sur le cancer, CHU Pontchaillou [Rennes], Service de Recherche en Hémato-Immunologie (SRHI - UMR_E 05), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris Diderot - Paris 7 (UPD7), Department of Molecular Cell Biology [Rehovot], Weizmann Institute of Science [Rehovot, Israël], Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-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)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Université Paris Diderot - Paris 7 (UPD7)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie-Institut National de la Santé et de la Recherche Médicale (INSERM), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), and Weizmann Institute of Science

Subjects
Small interfering RNA, Time Factors, Transcription, Genetic, [SDV]Life Sciences [q-bio], Apoptosis, Cell Cycle Proteins, medicine.disease_cause, S Phase, Mice, 0302 clinical medicine, Transcription (biology), Tumor Cells, Cultured, Cloning, Molecular, Phosphorylation, RNA, Small Interfering, Luciferases, Promoter Regions, Genetic, Glutathione Transferase, Oncogene Proteins, 0303 health sciences, Multidisciplinary, Kinase, Cell Cycle, Cell cycle, Protein-Tyrosine Kinases, Biological Sciences, Flow Cytometry, Proto-Oncogene Proteins c-bcl-2, 030220 oncology & carcinogenesis, Oxidoreductases, Protein Binding, G2 Phase, DNA, Complementary, Recombinant Fusion Proteins, Blotting, Western, Molecular Sequence Data, Mitosis, [SDV.CAN]Life Sciences [q-bio]/Cancer, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Protein Serine-Threonine Kinases, Gene product, 03 medical and health sciences, Complementary DNA, Proto-Oncogene Proteins, Two-Hybrid System Techniques, medicine, Animals, Humans, Amino Acid Sequence, RNA, Messenger, 030304 developmental biology, [SDV.GEN]Life Sciences [q-bio]/Genetics, Sequence Homology, Amino Acid, Tumor Suppressor Proteins, G1 Phase, Membrane Proteins, Oligonucleotides, Antisense, Blotting, Northern, Molecular biology, Precipitin Tests, Protein Structure, Tertiary, Protein Biosynthesis, Tumor Suppressor Protein p53, Carcinogenesis, HeLa Cells
Abstract

The p53 tumor suppressor protein plays a crucial role in tumorigenesis by controlling cell-cycle progression and apoptosis. We have previously described a transcript designated tumor suppressor activated pathway-6 (TSAP6) that is up-regulated in the p53-inducible cell line, LTR6. Cloning of the murine and human full-length TSAP6 cDNA revealed that it encodes a 488-aa protein with five to six transmembrane domains. This gene is the murine and human homologue of the recently published rat pHyde. Antibodies raised against murine and human TSAP6 recognize a 50- to 55-kDa band induced by p53. Analysis of the TSAP6 promoter identified a functional p53-responsive element. Functional studies demonstrated that TSAP6 antisense cDNA diminished levels of the 50- to 55-kDa protein and decreased significantly the levels of p53-induced apoptosis. Furthermore, TSAP6 small interfering RNA inhibited apoptosis in TSAP6-overexpressing cells. Yeast two-hybrid analysis followed by GST/ in vitro -transcribed/translated pull-down assays and in vivo coimmunoprecipitations revealed that TSAP6 associated with Nix, a proapoptotic Bcl-2-related protein and the Myt1 kinase, a negative regulator of the G 2 /M transition. Moreover, TSAP6 enhanced the susceptibility of cells to apoptosis and cooperated with Nix to exacerbate this effect. Cell-cycle studies indicated that TSAP6 could augment Myt1 activity. Overall, these data suggest that TSAP6 may act downstream to p53 to interface apoptosis and cell-cycle progression.

Published
2003
Full Text
View/download PDF

321. Asparagine Endopeptidase Controls Anti-Influenza Virus Immune Responses through TLR7 Activation

Author

Signe Hässler, Mira Tohme, Fany Blanc, Delphyne Descamps, Sophia Maschalidi, Fernando E. Sepulveda, Mustapha Si-Tahar, Peter van Endert, Michel Chignard, Bénédicte Manoury, ProdInra, Migration, Maladies Infectieuses et environnement - Etude des mécanismes d'activation et rôle des lymphocytes T Natural Killer lors des infections virales - - NKTVIR2008 - ANR-08-MIEN-0021 - MIE - VALID, U 1013, Institut National de la Santé et de la Recherche Médicale (INSERM), Faculté de Médecine, Université de Ngozi, Défense innée et inflammation, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Pathologies Respiratoires : Protéolyse et Aérosolthérapie, Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Francois Rabelais [Tours], INSERM (ANR) 008 01, ARC, Institut Curie Ph.D. fellowship, This study was supported by grants from INSERM (ANR 2010 MIDI 008 01 to B. M and ANR 2008 NKTVir to M. S), an ARC PhD fellowship to S. M, an Institut Curie Ph.D. fellowship to M. T, FRM and ANR post-doc support respectively to S. H and D. D., ANR-08-MIEN-0021,NKTVIR,Etude des mécanismes d'activation et rôle des lymphocytes T Natural Killer lors des infections virales(2008), Manoury, Bénédicte, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Male, RNA viruses, [SDV]Life Sciences [q-bio], Immunofluorescence, Antigen Processing and Recognition, CD8-Positive T-Lymphocytes, Adaptive Immunity, medicine.disease_cause, Mice, Influenza A Virus, H1N1 Subtype, 0302 clinical medicine, Bone Marrow, Viral classification, Influenza A virus, Lymphoid Organs, Biology (General), Receptor, Immune Response, Mice, Knockout, 0303 health sciences, Membrane Glycoproteins, T Cells, virus diseases, Cross-presentation, Immunizations, Innate Immunity, 3. Good health, [SDV] Life Sciences [q-bio], Cytokines, Signal transduction, Signal Transduction, Research Article, QH301-705.5, Immune Cells, Immunology, Antigen-Presenting Cells, Immunopathology, Biology, Microbiology, Virus, Immune Activation, Immune Deficiency, 03 medical and health sciences, Immune system, Orthomyxoviridae Infections, Antigen, Virology, Endopeptidases, Genetics, medicine, Animals, Immunoassays, Immunity to Infections, Molecular Biology, 030304 developmental biology, Inflammation, Host Cells, Immunity, Immune Defense, TLR7, RC581-607, Animal Models of Infection, Toll-Like Receptor 7, Immune System, Virulence Factors and Mechanisms, Immunologic Techniques, Parasitology, Immunologic diseases. Allergy, Viral Transmission and Infection, 030215 immunology
Abstract

Intracellular Toll-like receptors (TLRs) expressed by dendritic cells recognize nucleic acids derived from pathogens and play an important role in the immune responses against the influenza virus (IAV), a single-stranded RNA sensed by different receptors including TLR7. However, the importance of TLR7 processing in the development of anti-viral immune responses is not known. Here we report that asparagine endopeptidase (AEP) deficient mice are unable to generate a strong anti-IAV response, as demonstrated by reduced inflammation, cross presentation of cell-associated antigens and priming of CD8+ T cells following TLR7-dependent pulmonary infection induced by IAV. Moreover, AEP deficient lung epithelial- or myeloid-cells exhibit impaired TLR7 signaling due to defective processing of this receptor. Indeed, TLR7 requires a proteolytic cleavage by AEP to generate a C-terminal fragment competent for signaling. Thus, AEP activity is critical for TLR7 processing, opening new possibilities for the treatment of influenza and TLR7-dependent inflammatory diseases., Author Summary Influenza A virus, a negative stranded RNA, can cause severe illness in humans and animals and stimulates many receptors including Toll like receptors 7 (TLR7). TLR signaling induces maturation of dendritic cells and the production of a variety of inflammatory cytokines that are crucial for both innate and adaptive immunity. TLR7 is an intracellular receptor, which resides in endosomes and senses viruses to trigger host defence. Previous data have shown that TLR9 requires proteolysis to be functional but it is unclear whether other intracellular TLRs (TLR3 and TLR7) are also subject to degradation. Here, we used a protease deficient mouse model to show the in vivo importance of TLR7 processing in influenza infection. Inflammation monitored by cytokine release and adaptive immunity measured by cross priming of CD8+ T cells was significantly reduced in infected protease-deficient animals in comparison to control animals. We showed that TLR7 requires a proteolytic cleavage by a cysteine endopeptidase in order to be functional. Our findings indicate that TLR7 processing mediated by a protease, asparagynil endopeptidase, is critical for inducing robust anti-influenza immune responses. Given our results, targeting TLR7 response in the lungs through proteases may offer new therapeutic potential in pulmonary infection.

Published
2012
Full Text
View/download PDF

322. Antigen Recognition By Autoreactive Cd4+ Thymocytes Drives Homeostasis Of The Thymic Medulla

Author

Beat A. Imhof, Lucia Guerri, Jeanne Guenot, Adrian Liston, Ed Palmer, Olivier Lantz, Arnauld Sergé, Walter Reith, Magali Irla, Sergé, Arnauld, Université de Genève = University of Geneva (UNIGE), Immunité et cancer (U932), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), and University Hospital Basel [Basel]

Subjects
Male, Anatomy and Physiology, [SDV]Life Sciences [q-bio], Immunofluorescence, CD40 Ligand/metabolism, Gene Expression, lcsh:Medicine, Autoimmunity, Adaptive Immunity, ddc:616.07, Autoantigens, Tissue Culture Techniques, Mice, 0302 clinical medicine, Immune Physiology, Homeostasis, Lymphoid Organs, lcsh:Science, Lymphotoxin-alpha, Antigens, CD40/metabolism, Mice, Knockout, 0303 health sciences, Thymocytes, Multidisciplinary, Receptor Activator of Nuclear Factor-kappa B, hemic and immune systems, Cell biology, Thymocytes/immunology/metabolism/physiology, [SDV] Life Sciences [q-bio], CD4 Antigens, Cytokines, Medicine, Female, Lymphotoxin-alpha/genetics/metabolism, Epithelial Cells/metabolism/physiology, Central tolerance, Signal transduction, Research Article, Signal Transduction, Lymphotoxin alpha, Thymus Gland/cytology/growth & development/immunology, CD40 Ligand, Immunology, Antigens, CD4/metabolism, chemical and pharmacologic phenomena, Thymus Gland, Biology, Lymphotoxin beta Receptor/metabolism, 03 medical and health sciences, Lymphotoxin beta Receptor, Autoantigens/immunology, Immune Tolerance, Animals, CD40 Antigens, Medulla, Body Patterning, Cell Proliferation, 030304 developmental biology, CD86, T-cell receptor, lcsh:R, Immunity, Epithelial Cells, Mice, Inbred C57BL, Lymphotoxin, Immune System, Immunologic Techniques, Clinical Immunology, lcsh:Q, Receptor Activator of Nuclear Factor-kappa B/metabolism, Lymphotoxin beta receptor, 030215 immunology
Abstract

International audience; The thymic medulla is dedicated for purging the T-cell receptor (TCR) repertoire of self-reactive specificities. Medullary thymic epithelial cells (mTECs) play a pivotal role in this process because they express numerous peripheral tissue-restricted self-antigens. Although it is well known that medulla formation depends on the development of single-positive (SP) thymocytes, the mechanisms underlying this requirement are incompletely understood. We demonstrate here that conventional SP CD4⁺ thymocytes bearing autoreactive TCRs drive a homeostatic process that fine-tunes medullary plasticity in adult mice by governing the expansion and patterning of the medulla. This process exhibits strict dependence on TCR-reactivity with self-antigens expressed by mTECs, as well as engagement of the CD28-CD80/CD86 costimulatory axis. These interactions induce the expression of lymphotoxin α in autoreactive CD4⁺ thymocytes and RANK in mTECs. Lymphotoxin in turn drives mTEC development in synergy with RANKL and CD40L. Our results show that Ag-dependent interactions between autoreactive CD4⁺ thymocytes and mTECs fine-tune homeostasis of the medulla by completing the signaling axes implicated in mTEC expansion and medullary organization.

Published
2012
Full Text
View/download PDF

323. Pertussis toxin reduces the number of splenic Foxp3(+) regulatory T cells

Author

Roland S. Liblau, Lennart T. Mars, Nicolas Couturier, Eliane Piaggio, Daniel Gonzalez-Dunia, Cécile Cassan, Sabine Desbois, Jacques P. Zappulla, Jan Bauer, Florence Bucciarelli, Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud]), Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de recherches de biochimie macromoléculaire (CRBM), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-IFR122-Centre National de la Recherche Scientifique (CNRS), Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Pôle neurosciences [Hôpital de Purpan - Toulouse], CHU Toulouse [Toulouse], Department Neuroimmunology, Medizinische Universität Wien = Medical University of Vienna-Center for Brain Research, Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Pôle Neurosciences [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Medizinische Universität Wien = Medical University of Vienna-Center for Brain Research [Vienna, Austria], and Medizinische Universität Wien = Medical University of Vienna

Subjects
Lymphocyte, [SDV]Life Sciences [q-bio], experimental autoimmune encephalomyelitis, PI, medicine.disease_cause, T-Lymphocytes, Regulatory, Autoimmunity, Mice, cites 73 articles This article PTx, 0302 clinical medicine, T-Lymphocyte Subsets, Immunology and Allergy, IL-2 receptor, ComputingMilieux_MISCELLANEOUS, propidium iodide, 0303 health sciences, EAE, Experimental autoimmune encephalomyelitis, Antibodies, Monoclonal, FOXP3, Drug Synergism, Forkhead Transcription Factors, hemic and immune systems, Growth Inhibitors, regulatory T, mean fluorescence intensity, 3. Good health, Treg, medicine.anatomical_structure, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, medicine.symptom, Immunosuppressive Agents, MFI, Immunology, chemical and pharmacologic phenomena, Inflammation, Biology, Pertussis toxin, Lymphocyte Depletion, 03 medical and health sciences, Immune system, medicine, Animals, 030304 developmental biology, pertussis toxin, Receptors, Interleukin-2, medicine.disease, Mice, Inbred C57BL, Spleen, 030215 immunology
Abstract

Pertussis toxin (PTx) is a bacterial toxin used to enhance the severity of experimental autoimmune diseases such as experimental autoimmune encephalomyelitis. It is known to promote permeabilization of the blood-brain barrier, maturation of APC, activation of autoreactive lymphocytes and alteration of lymphocyte migration. In this study, we show that i.v. injection of PTx in mice induces a decrease in the number of splenic CD4+CD25+ regulatory T cells (Treg cells). Furthermore, PTx not only induces a depletion of the dominant CD4+CD25+Foxp3+ subpopulation of splenic Treg cells, but also reduces to a similar extent the CD4+CD25−Foxp3+ subpopulation. On a per cell basis, the suppressive properties of the remaining Treg cells are not modified by PTx treatment. The reduction in splenic Treg cells is associated with preferential migration of these cells to the liver. Additionally, Treg cells exhibit a high sensitivity to PTx-mediated apoptosis in vitro. Finally, in vivo depletion of Treg cells by injection of an anti-CD25 Ab, and PTx treatment, present synergistic experimental autoimmune encephalomyelitis exacerbating effects. Therefore, we identify a new effect of PTx and provide an additional illustration of the influence of microbial components on the immune system affecting the balance between tolerance, inflammation and autoimmunity.

Full Text
View/download PDF

324. Role of MR1-driven signals and amphiregulin on the recruitment and repair function of MAIT cells during skin wound healing

Author

Anastasia du Halgouet, Aurélie Darbois, Mansour Alkobtawi, Martin Mestdagh, Aurélia Alphonse, Virginie Premel, Thomas Yvorra, Ludovic Colombeau, Raphaël Rodriguez, Dietmar Zaiss, Yara El Morr, Hélène Bugaut, François Legoux, Laetitia Perrin, Selim Aractingi, Rachel Golub, Olivier Lantz, Marion Salou, Immunité et cancer (U932), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Chimie biologique des membranes et ciblage thérapeutique (CBMCT - UMR 3666 / U1143), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Universität Regensburg (UR), University Hospital Regensburg, Leibniz Institute of Immunotherapy - University of Regensburg, Lymphocytes et Immunité - Lymphocytes and Immunity, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Institut Curie [Paris], Centre d'Investigation Clinique en Biotherapie des cancers (CIC 1428 , CBT 507 ), Institut Gustave Roussy (IGR)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), This study received funding from the following institutions: Institut National de la Santé et de la Recherche Médicale (O.L.), Institut Curie (O.L.), Agence Nationale de la Recherche Grant MAITANR-16-CE15-0020-01 (O.L.), Agence Nationale de la Recherche Grant MAIT-repair ANR-20-CE15-0028-01 (O.L.), Agence Nationale de la Recherche ANR-10-IDEX-0001-02 PSL (O.L.), European Research Council ERC-2019-ADG-885435 (O.L.), and Fondation pour la Recherche Médicale FDT202106013036 (A.d.H.)., ANR-16-CE15-0020,MAIT,Biologie des cellules MAIT in vivo en situation normale ou pathologique(2016), ANR-20-CE15-0028,MAIT-repair,Mécanisme de la fonction de réparation tissulaire des lymphocytes MAIT(2020), and ANR-10-IDEX-0001,PSL,Paris Sciences et Lettres(2010)

Subjects
MESH: Minor Histocompatibility Antigens, MESH: Humans, [SDV]Life Sciences [q-bio], Immunology, MAIT cells, MESH: Receptors, Antigen, T-Cell, MESH: Amphiregulin, MESH: Mucosal-Associated Invariant T Cells, CXCR6 chemokine receptor, MESH: Histocompatibility Antigens Class I, TCR signaling, MESH: Wound Healing, Infectious Diseases, scRNA-seq, Immunology and Allergy, MESH: Animals, amphiregulin, tissue repair, skin excision, MESH: Mice
Abstract

International audience; Tissue repair processes maintain proper organ function following mechanical or infection-related damage. In addition to antibacterial properties, mucosal associated invariant T (MAIT) cells express a tissue repair transcriptomic program and promote skin wound healing when expanded. Herein, we use a human-like mouse model of full-thickness skin excision to assess the underlying mechanisms of MAIT cell tissue repair function. Single-cell RNA sequencing analysis suggested that skin MAIT cells already express a repair program at steady state. Following skin excision, MAIT cells promoted keratinocyte proliferation, thereby accelerating healing. Using skin grafts, parabiosis, and adoptive transfer experiments, we show that MAIT cells migrated into the wound in a T cell receptor (TCR)-independent but CXCR6 chemokine receptor-dependent manner. Amphiregulin secreted by MAIT cells following excision promoted wound healing. Expression of the repair function was probably independent of sustained TCR stimulation. Overall, our study provides mechanistic insights into MAIT cell wound healing function in the skin.

Full Text
View/download PDF

325. Macropinocytosis Overcomes Directional Bias in Dendritic Cells Due to Hydraulic Resistance and Facilitates Space Exploration

Author

Matthieu Piel, Ana-Maria Lennon-Duménil, Raphaël Voituriez, Maria-Graciela Delgado, Hélène D. Moreau, Jean-François Joanny, Odile Malbec, Zahraa Alraies, Carles Blanch-Mercader, Mathieu Maurin, Philippe Bousso, Doriane Sanséau, Rafaele Attia, Immunité et cancer (U932), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire Physico-Chimie Curie [Institut Curie] (PCC), Institut Curie [Paris]-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Compartimentation et dynamique cellulaires (CDC), Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Dynamiques des Réponses immunes - Dynamics of Immune Responses, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL), Chaire Matière molle et biophysique, Collège de France (CdF (institution)), Laboratoire Jean Perrin (LJP), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), Sorbonne Université (SU)-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), Institut Pierre-Gilles de Gennes pour la Microfluidique, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Collège de France - Chaire Matière molle et biophysique, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-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), and CCSD, Accord Elsevier

Subjects
Male, cell migration, dendritic cell, macropinocytosis, [SDV]Life Sciences [q-bio], Cell, microfluidics, Down-Regulation, barotaxis, Biology, Hydraulic resistance, General Biochemistry, Genetics and Molecular Biology, Cell Line, [PHYS] Physics [physics], Mice, 03 medical and health sciences, 0302 clinical medicine, Immune system, Cell Movement, medicine, Animals, Cytoskeleton, Molecular Biology, 030304 developmental biology, [PHYS]Physics [physics], theoretical physics, 0303 health sciences, Pinocytosis, Cell migration, Actomyosin, Dendritic Cells, Cell Biology, Dendritic cell, Fluid transport, microenvironment, Cell biology, Mice, Inbred C57BL, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, Hydrodynamics, Female, intravital imaging, 030217 neurology & neurosurgery, Developmental Biology
Abstract

International audience; The migration of immune cells can be guided by physical cues imposed by the environment, such as geometry, rigidity, or hydraulic resistance (HR). Neutrophils preferentially follow paths of least HR in vitro, a phenomenon known as barotaxis. The mechanisms and physiological relevance of barotaxis remain unclear. We show that barotaxis results from the amplification of a small force imbalance by the actomyosin cytoskeleton, resulting in biased directional choices. In immature dendritic cells (DCs), actomyosin is recruited to the cell front to build macropinosomes. These cells are therefore insensitive to HR, as macropinocytosis allows fluid transport across these cells. This may enhance their space exploration capacity in vivo. Conversely, mature DCs down-regulate macropinocytosis and are thus barotactic. Modeling suggests that HR may help guide these cells to lymph nodes where they initiate immune responses. Hence, DCs can either overcome or capitalize on the physical obstacles they encounter, helping their immune-surveillance function.

Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results