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Book
Identification des épitopes reconnus par des anticorps inhibiteurs de l'activation du TGF-ß1
Authors: --- ---
Year: 2015 Publisher: Bruxelles: UCL. Faculté de pharmacie et des sciences biomédicales,

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Regulatory T cells are involved in the inhibition of anti-tumor immune responses, in cancer patients. Tregs exert their immunosuppressive function through the production of active TGFß1.TGF-ß 1 is a potent immunosuppressive cytokine. It is produced by many cell types in an inactive form called latent TGF-BI , which needs to be activated to acquire its biological activity. In Tregs, TGF-ß 1 activation depends on GARP. This transmembrane protein binds and presents latent- TGF-ß 1 on the Treg surface. Our lab developed two monoclonal antibodies (MHG-8 and LHG-10) that bind GARP/TGF-BI complexes and inhibit TGF-ß 1 activation by human Tregs. The three dimensional structure of the Fab fragment of MHG-8 bound to GARPITGF- 1 complex has recently been solved by crystallography and X-ray diffraction. This allowed the identification of amino acids (aa) in GARP and TGF-ß 1 molecules that contact aa in the Fab of the MHG-8.The aim of my project was to identify, among the 36 aa of the GARP/ TGF-ß 1 complex that contact the Fab of the MHG-8 , those that are important for the binding and for the inhibitory function. 1 constructed and transfected plasmids coding for mutated forms of GARP and TGF-ß 1 in 293T cells. J analyzed the binding of MHG-8 and LHG 10 to the mutant proteins by flow cytometry. 1identified 5 aa in GARP and 1 in TGF-ß 1 that are important for the binding of MHG-8, and 2 aa in GARP and 1 aa in TGF-ß 1, that are important for the binding of LHG-10. 1 also set up a functional test in transfected cells, to measure the ability of MHG-8 and LHG-10 to inhibit TGF­ activation from GARP/ TGF-ß 1 complexes containing mutated forms of GARP or TGF-ß 1. Among the aa shown to be non-essential for the binding of the antibodies, 1 identified 2 aa in GARP that, when mutated, affect the inhibitory function of MHG-8 and one in TGF-ß 1 that when mutated affects the inhibitory function of LHG-10.This work may help to better define the molecular mechanisms by which anti-GARP antibodies with therapeutic potential inhibit the immunosuppressive function of Tregs. Les lymphocites T régulateurs (Tregs) contribuent à l’inhibition des réponses immunitaires dirigées contre les tumeurs chez les patients cancéreux. Les Tregs exercent leur action immunosuppressive au moins en partie via la production de TGF-β1 actif. Le TGF-β1 est une cytokine immunosuppressive puissante. Il est produit par de nombreux types cellulaires sous une forme latente, inactive. L’activation du TGF-β1 latent par les tregs dépend de la protéine transmembranaire GARP, qui lie et présente le TGF-β1 latent à la surface des tregs. Le laboratoire a développé deux anticorps monoclonaux (MHG-8 et LHG10) qui lient les complexes GARP/ TGF-β1 et inhibent l’activation du TGF-β1 par les tregs humains. La structure tridimensionnelle du fragment Fab du MHG-8 associé au complexe GARP/ TGF-β1 a récemment été résolue par cristallographie et diffraction aux rayons X. Ceci a permis d’identifier les acides aminés (aa) de GARP et du TGF-β1 qui sont en contact avec des aa du Fab du MHG-8. L’objectif de mon travail, était d’identifier, parmi les 36 aa du complexe GARP/ TGF-β1 qui sont en contact avec le Fab du MHG-8, ceux qui sont importants pour la liaison et pour l’activité inhibitrice des anticorps MHG-8 et du LHG-10. J’ai construit et transféré des plasmides codant pour des formes mutées de GARP et du TGF-β1 dans des cellules 293T, et j’ai analysé la liaison du MHG-8 et du LHG-10 aux cellules transfectées par cytométrie en flux. Ceci m’a permis d’identifier 5 aa de GARP et 1 aa du TGF-β1 importants pour la liaison du MHG-8, ainsi que 2 aa de GARP et 1 aa du TGF-β1 importants pour la liaison du LHG-10. J’ai également mis au point un test fonctionnel dans des cellules transfectées, me permettant de déterminer la capacité du MHG-8 et du LHG-10 à inhiber la production de TGF-β1 actif à partir de complexes GARP/ TGF-β1 contenant une forme mutée de GARP ou du TGF-β1. Parmi les aa qui ne sont pas essentiels pour la liaison aux, anticorps, j’ai identifié 2aa de GARP dont la mutation affecte la capacité inhibitrice du MHG-8, et 1 aa du TGF-β1 dont la mutation affecte la capacité inhibitrice du LHG-10. Mon travail pourrait servir de base pour mieux comprendre les mécanismes moléculaires par lesquels des anticorps anti-GARP possédant un potentiel de développement clinique inhibent la fonction immunosuppressive des tregs.


Book
Etude de la superfamille du TGFβ durant le développement de la thyroïde
Authors: --- ---
Year: 2012 Publisher: Bruxelles: UCL,

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Book
Advances in Mechanisms of Renal Fibrosis
Authors: ---
Year: 2018 Publisher: Frontiers Media SA

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Scarring of the glomerular and tubulointerstitial compartments is a hallmark of progressive kidney disease. Renal fibrosis involves a complex interplay between kidney cells, leukocytes and fibroblasts in which transforming growth factor-β (TGF-β) plays a key role. This eBook provides a comprehensive update on TGF-β signalling pathways and introduces a range of cellular and molecular mechanisms involved in renal fibrosis both upstream and downstream of TGF-β. The wide variety of potential new targets described herein bodes well for the future development of effective therapies to tackle the major clinical problem of progressive renal fibrosis.

Keywords

TGF-beta --- fibroblast --- JNK --- miRNA --- Smad --- non-classical RAS --- BMP7 --- HDAC --- HIPK2


Book
Advances in Mechanisms of Renal Fibrosis
Authors: ---
Year: 2018 Publisher: Frontiers Media SA

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Abstract

Scarring of the glomerular and tubulointerstitial compartments is a hallmark of progressive kidney disease. Renal fibrosis involves a complex interplay between kidney cells, leukocytes and fibroblasts in which transforming growth factor-β (TGF-β) plays a key role. This eBook provides a comprehensive update on TGF-β signalling pathways and introduces a range of cellular and molecular mechanisms involved in renal fibrosis both upstream and downstream of TGF-β. The wide variety of potential new targets described herein bodes well for the future development of effective therapies to tackle the major clinical problem of progressive renal fibrosis.

Keywords

TGF-beta --- fibroblast --- JNK --- miRNA --- Smad --- non-classical RAS --- BMP7 --- HDAC --- HIPK2


Book
Advances in Mechanisms of Renal Fibrosis
Authors: ---
Year: 2018 Publisher: Frontiers Media SA

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Abstract

Scarring of the glomerular and tubulointerstitial compartments is a hallmark of progressive kidney disease. Renal fibrosis involves a complex interplay between kidney cells, leukocytes and fibroblasts in which transforming growth factor-β (TGF-β) plays a key role. This eBook provides a comprehensive update on TGF-β signalling pathways and introduces a range of cellular and molecular mechanisms involved in renal fibrosis both upstream and downstream of TGF-β. The wide variety of potential new targets described herein bodes well for the future development of effective therapies to tackle the major clinical problem of progressive renal fibrosis.

Keywords

TGF-beta --- fibroblast --- JNK --- miRNA --- Smad --- non-classical RAS --- BMP7 --- HDAC --- HIPK2

Transforming Growth Factor-Beta Protocols
Author:
ISBN: 0896036464 9786610820450 1280820454 1592590535 Year: 2000 Publisher: Totowa, NJ : Humana Press : Imprint: Humana,

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The recent isolation and identification of the TGFß receptors, as well as their intracellular mediating proteins, has driven the development of many new methodologies and techniques for the study of TGFß. In Transforming Growth Factor-Beta Protocols, Philip H. Howe and a group of well-versed experimentalists present the first major collection of indispensable classic and cutting-edge TGFß assays. Described in great detail to ensure robust and successful results, these readily reproducible techniques range from the growth inhibition assay for TGFß to methodologies for monitoring its interactions with the mediating proteins. Extensive notes discuss potential pitfalls and provide tips on how to avoid failures, and throughout, emphasis is given to detailing those technical steps critical for experimental success that are often omitted in the primary literature. Concise and highly practical, Transforming Growth Factor-Beta Protocols provides today's molecular and cell biologists-both expert and novice-with time-tested methods for the identification and analysis of the signal transduction pathways by which TGFß induces and modulates physiological behavior.

Transforming growth factor-betas : chemistry, biology and therapeutics
Authors: ---
ISBN: 0897665732 0897665740 Year: 1990 Volume: 593 Publisher: New York, NY : New York Academy of Sciences,


Book
TGF-SS in human disease
Authors: ---
ISBN: 4431544089 4431544097 Year: 2013 Publisher: Tokyo ; New York : Springer,

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Transforming growth factor-β (TGF-β) is a secreted polypeptide with multifunctional properties manifested during embryonic development, adult organ physiology, and pathobiology of major diseases, including cancer and fibrotic and cardiovascular diseases. The signaling pathway of TGF-β now is rather well understood. Continuing revelations in the mechanisms of action of TGF-β provide specific mechanistic examples of how human cells lose their controlled function and behave wrongly during the development of diverse diseases. Equally important, however, is the current promise of exploiting the TGF-β pathway in combating human disease. This book comprehensively covers major areas of human disease where the involvement of TGF-β is firmly established. Simultaneously, the book highlights major gaps in knowledge and the future directions of research that can benefit human medical science. The core set of diseases where TGF-β action is well documented and are included in the book are cancer and cardiovascular and fibrotic disorders. The central aim of the book is to stimulate young scientists to enter the prolific TGF-β field and find new solutions to the problems remaining in this area of study. For this purpose the book provides authoritative educational chapters that furnish a good introduction to the field for young doctoral students, postdocs, and clinical fellows. The book also serves as a valuable reference for the aficionados in the field, who can find accessible and well-illustrated material for their teaching and lecturing activities, via which the importance of TGF-β biology is disseminated to the world of science and to the public.


Book
Aging, neurogenesis and neuroinflammation in hearing loss and protection
Authors: --- --- ---
ISBN: 2889196445 9782889196449 Year: 2015 Publisher: Frontiers Media SA

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Worldwide, 278 million people are estimated to have moderate to profound hearing loss. Age-related hearing loss, also known as presbyacusis, affects approximately half of the population over 60 years old, making it the second most common cause of disability in older people. Hearing loss occurs when the sensory cells and neurons of the cochlea degenerate and die. The vestibular system, which holds the sense of balance, shares a common embryonic origin with the cochlea and together conform the inner ear. Balance problems are a trait of ageing to the point that balance ability is considered a sensor of physical decline and vestibular degeneration is the most common cause of falls in the elderly. Still the molecular bases of ageing in the vestibular system have not been studied in detail. Genetic and environmental factors contribute to the progression of age-related hearing loss (ARHL). Being noise the main environmental noxious agent for human hearing in the industrialized societies.There is no restorative treatment for deafness but functional replacement by means of prosthesis. Therefore, prevention and treatment of hearing loss is an unmet medical need. To develop innovative medical strategies against hearing loss, it is critical to understand the causes of ARHL and the essential pathways responsible for the manifestation of this complex disease.In this research topic, experts will discuss the stages and molecular elements of the damage and repair processes involved in ARHL, from cellular processes involved in ageing as senescence and autophagy, to molecules essential for hearing as IGF-1 and neurotrophins. Neuroinflammation takes a central stage as an essential element in the progression of injury and cell loss, and a target for cell protection strategies. Neurogenesis is also essential to understand the adult cochlea self-repair potential. Finally, the mechanisms of action and the potential of novel therapies for hair cell repair and protection will be discussed along with drug delivery strategies.


Book
Stem Cell and Biologic Scaffold Engineering
Author:
ISBN: 3039214985 3039214977 Year: 2019 Publisher: MDPI - Multidisciplinary Digital Publishing Institute

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Tissue engineering and regenerative medicine is a rapidly evolving research field which effectively combines stem cells and biologic scaffolds in order to replace damaged tissues. Biologic scaffolds can be produced through the removal of resident cellular populations using several tissue engineering approaches, such as the decellularization method. Indeed, the decellularization method aims to develop a cell-free biologic scaffold while keeping the extracellular matrix (ECM) intact. Furthermore, biologic scaffolds have been investigated for their in vitro potential for whole organ development. Currently, clinical products composed of decellularized matrices, such as pericardium, urinary bladder, small intestine, heart valves, nerve conduits, trachea, and vessels, are being evaluated for use in human clinical trials. Tissue engineering strategies require the interaction of biologic scaffolds with cellular populations. Among them, stem cells are characterized by unlimited cell division, self-renewal, and differentiation potential, distinguishing themselves as a frontline source for the repopulation of decellularized matrices and scaffolds. Under this scheme, stem cells can be isolated from patients, expanded under good manufacturing practices (GMPs), used for the repopulation of biologic scaffolds and, finally, returned to the patient. The interaction between scaffolds and stem cells is thought to be crucial for their infiltration, adhesion, and differentiation into specific cell types. In addition, biomedical devices such as bioreactors contribute to the uniform repopulation of scaffolds. Until now, remarkable efforts have been made by the scientific society in order to establish the proper repopulation conditions of decellularized matrices and scaffolds. However, parameters such as stem cell number, in vitro cultivation conditions, and specific growth media composition need further evaluation. The ultimate goal is the development of “artificial” tissues similar to native ones, which is achieved by properly combining stem cells and biologic scaffolds and thus bringing them one step closer to personalized medicine. The original research articles and comprehensive reviews in this Special Issue deal with the use of stem cells and biologic scaffolds that utilize state-of-the-art tissue engineering and regenerative medicine approaches.

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