Déficits congénitaux de l'immunité cellulaire et thérapie cellulaire : l'apport de la thérapie cellulaire dans la prévention des complications survenant au décours des allogreffes de cellules souches hématopoïétiques

Allogeneic hematopoietk stem cell transplantation (HSCT) is the only way to cure severe Primary Immune Deficiencies (PID). However, HSCT is, sometimes complicated by infections or Graft vers us Host disease (GVHD) which can lead to patient death. In this work we illustrate how, for three PIDs, Hyper IgM syndrome (HIGM), Wiskott Aldrich Syndrome (WAS) and DiGeorge syndrome (cDGS), expert knowledge of the disease and its complications together with optimal management of the allogeneic HSCT procedure both increase the chances for a successful outcome. Analysing post-HSCT infections, we explain how B-cell depletion is very effective in preventing EBV -related lymphoproliferative syndromes. We also report the successful treatment of a pre-transplant Cryptosporidium parvum infection in two children with X-linked HIGM by an allogeneic HSCT that led to cure of the CD40L deficiency. Nevertheless, such an effective result seems to be favored by the absence of severe hepatic impairment at the time of transplant. We then study how to prevent GVHD by graft manipulations. We report CD34-positive selection limits in a patient with W AS and we also present different alloreactive depletion methods to prevent GVHD while preserving graft anti-infectious capacity. Finally, in the specifie case of cDGS, we report on the effectiveness of unmanipulated peripheral blood mononuclear cell infusion to avoid a conditioning regimen and to simplify allogeneic transplantation procedure.L'apport de la Thérapie cellulaire dans la prévention des complications survenant au décours des allogreffes de Cellules souches hématopoïétiques. L'allogreffe de cellules souches hématopoïétiques (CSH) est, à ce jour, le seul traitement curatif des déficits immunitaires (DI) héréditaires sévères. Cependant, les complications des allogreffes parmi lesquelles les infections et la réaction du greffon contre l'hôte sont parfois létales. Ce travail s'attache, à travers trois modèles de déficit immunitaire héréditaire - les syndromes hyper IgM (HIGM), le syndrome de Wiskott Aldrich (W AS) et le Syndrome de DiGeorge (cDGS) - à montrer qu'une bonne connaissance du DI et de ses complications d'une part ainsi que l'adaptation du protocole d'allogreffe accroissent les chances de succès de cette thérapeutique. Dans le cadre des infections survenant au décours des allogreffes de CSH, nous rapportons comment la déplétion en lymphocytes B du greffon permet de prévenir efficacement la survenue d'un lymphome EBV-induit après allogreffes. Puis, nous décrivons l'effet bénéfique de l'allogreffe de CSH sur une infection à Cryptosporidium parvum chez deux enfants atteints d'un HIGM lié à l'X. Il semble que cet effet ne soit obtenu que lorsque les atteintes hépatiques engendrées par ce parasite ne sont pas trop avancées. Nous étudions ensuite la prévention de la réaction du greffon contre l'hôte (GVHD) par manipulation du greffon de CSH. Nous rapportons les limites de la sélection positive CD34+ chez un patient atteint de W AS puis nous exposons les techniques de déplétion alloréactive permettant de conserver le capital anti-infectieux du greffon. Enfin, dans le contexte particulier du cDGS, nous montrons comment simplifier la procédure d'allogreffe en ne réinjectant que les cellules d'intérêt et en s'affranchissant du conditionnement et de ses effets secondaires

Reconstitution immunitaire et immunothérapie adoptive anti-virales après allogreffe de cellules souches hématopoiétiques

L allogreffe de cellules souches hématopoïétiques (CSH) est un traitement efficace des Hémopathies malignes. Cependant, les complications des allogreffes parmi lesquelles les infections virales sont associées parfois à une morbidité et une mortalité importantes. Ces infections surviennent en l absence de reconstitution immunitaire. Un monitoring régulier de la charge virale des principaux agents infectieux impliqués est réalisé mais amène parfois à la mise en oeuvre abusive de traitements anti-viraux qui ne sont pas dénués de toxicité.Dans ce travail, nous proposons d associer à ce monitoring un suivi régulier de la reconstitution immunitaire spécifique afin de cibler parmi les patients présentant une réactivation ceux qui nécessitent un traitement curatif de ceux qui pourront maîtriser l infection par leur système immunitaire. Nous illustrons ce propos avec le virus d Epstein Barr (EBV) et avons en cours une étude sur l Adénovirus (ADV).Dans certains cas parfaitement ciblés, les traitements anti-viraux s avèrent inefficaces. C est pourquoi dans ce travail, nous présentons la mise au point d une technique de grade clinique de production de lymphocytes T cytotoxiques anti-ADV (CTL anti-ADV) en condition GMP (Good Manufacturing Practice), grâce au système CliniMACS et au Cytokine Capture System de Miltenyi, afin de proposer une immunothérapie adoptive.Nous décrivons par la suite trois expériences cliniques de traitement compassionnel d une infection ADV post-allogreffe de CSH. Enfin, nous présentons les résultats préliminaires de la production de CTL bispécifique anti-ADV et CMVHematopoietic stem cells Transplantation (HSCT) is a well recognized strategy for treatment of haematological malignancies. However, HSCT complications among which the viral infections a reassociated with high morbidity and mortality. These infections arise in the absence of immune reconstitution. Monitoring of viral reactivations after allogeneic HSCT is necessary, to identify patients at risk of viral infections, but not sufficient, as patients may be abusively treated. In this work we propose to combine viral DNA load assessment with specific immune monitoring to target patients who need to be treated. We report a retrospective study investigating EBV infection and EBV-specific immune recovery using the functional IFN Elispot assay in 40 allogeneic HSCT patients. We initiated a similar study with ADV which is pending. However, although patients are correctly targeted, anti-viral treatment is sometimes not effective. We present a study on the development of a complete clinical grade generation of Human anti-Adenovirus cytotoxic T cells in GMP (Good Manufacturing Practice) conditions, thanks to the system CliniMACS and the Cytokine Capture System, to propose an adoptive immunotherapy to the recipient.We describe afterwards three clinical experiments of treatment of an ADV infection after HSCT.Finally, we present the preliminary results of the anti-ADV and -CMV bi-specific CTL production.NANCY1-Bib. numérique (543959902) / SudocSudocFranceF

IL-15 as a potential target in leukemia

International audienceLeukemia, one of the most aggressive hematopoietic malignancies, is characterized by excessive proliferation, survival, and impaired differentiation of hematopoietic stem cells. Interleukin 15, a proinflammatory cytokine, induces proliferation and promotes cell survival of human T and B lymphocytes, as well as natural killer cells. However, it may also play a detrimental role in the onset of leukemia. This review provided an overview of the aberrant expression of Interleukin 15 and its role in the development and progression of this hematological malignancy. Also, we critically explored the potential therapeutic opportunities involved in targeting the disruption of interleukin-15 signaling as well as in interleukin-15-mediated enhancement of antitumor immunity

Caractérisation des cellules souches mésenchymateuses du sang placentaire et de la gelée de Wharton

Les cellules souches suscitent de grands espoirs pour la thérapie cellulaire et l'ingénierie tissulaire. Les CSM du tissus foetaux (sang placentaire et gelée de Wharton du cordon ombilical), à l'origine d'épiblaste embryonnaire, sont considérées comme plus primitives que les CSM provenant de sources adultes. Les conditions de culture ayant un impact sur le comportement des cellules, dans notre étude, nous avons exploré l'effet de la concentration de l'oxygène sur l'expansion, l'immunophénotypage et la différenciation de ces cellules. L'objectif de ce travail est d'identifier la méthode optimale d'isolation des CSM issues de tissus foetaux. Compte tenu du faible taux de succès dans l'isolement des CSM extraites du sang placentaire, nous nous sommes dirigés vers les CSM-GW. Nous y avons déterminé in situ, les marqueurs spécifiques exprimés dans la gelée de Wharton et à la périphérie. Des études sur la morphologie, la cinétique de croissance, et sur l'expression phénotypique des marqueurs de surface, des CSM-GW, ont été effectuées sur une longue durée (7 passages) à différentes conditions de culture. Nous avons montré que la GW est composée d'une abondante matrice extracellulaire riche en collagènes et glycosaminoglycannes et que les cellules possèdent un phénotype variable selon leur localisation dans la gelée. Ce tissu est capable de fournir une quantité importante de CSM (6,7x105 Cs/cm de cordon) qui gardent une morphologie constante. Enfin, quel que soit le passage, la concentration de l'oxygène ne semble pas avoir d'effet sur le phénotype des cellules. En revanche, une faible teneur en oxygène durant l'expansion semble diminuer le temps de doublement des cellules, favoriser la chondrogénèse et inhiber la différenciation ostéogénique. Enfin, quelles que soient les conditions de culture, la différenciation adipogénique des CSM-GW semble difficile à obtenirStem cells are the hopes for cell therapy and tissue engineering. MSCs from fetal tissue (umbilical cord blood and WJ), which are a source of embryonic epiblast grow relatively faster comparing to other adult sources. The culture condition can affect cell behavior. In our study, we explored the effect of oxygen concentration on the expansion, immunophenotyping, and differentiation of these cells. The aim of this work is to identify the optimal method for isolation of MSCs derived from fetal tissue. Given the low rate of success in the isolation of MSCs from cord blood, we headed to WJ-MSCs. We have determined in siu, the specific markers expressed in the WJ and in the perivascular region. Studies on the morphology growth kinetics, and phenotypic expression of surface makers of MSCs isolated from WJ were made over a long period (7 passages) in different culture conditions. We have shown that WJ is composed of an abundant extracellular matrix rich in collagen and glycasominoglycans and have variable phenotype depending from their localization in the jelly. This tissue is able to provide a large amount of MSCs (6.7x105 Cs/cm of cord) that maintain a constant morphology. Finally, regardless of the passage, the oxygen concentration does not effect on the phenotype of the cells. In contrast, a low oxygen concentration during expansion appears to decrease the doubling time of MSCs, promote chondrogenesis and inhibit osteogenic differentiation. Finally, whatever the culture conditions, adipogenic differentiation of WJ-MSC seems difficult to obtainMETZ-SCD (574632105) / SudocNANCY1-Bib. numérique (543959902) / SudocNANCY2-Bibliotheque electronique (543959901) / SudocNANCY-INPL-Bib. électronique (545479901) / SudocSudocFranceF

Mécanotransduction et cellules souches mésenchymateuses humaines (Etude de l'effet d'une contrainte d'étirement équiibiaxial sur la voie de TGF-ß)

Le TGF-ß (Transforming growth factor-ß) joue un rôle très important dans la différenciation des cellules souches mésenchymateuses humaines (CSMs) en chondrocytes. La contrainte mécanique joue un rôle important aussi dans la différenciation des CSMs, mais, le mécanisme de mécanotransduction reste obscur. Le but de nos travaux a été de définir l effet; a des temps courts de l étirement équibiaxial (0,5 Hz ou 1 Hz, 5%) sur les CSMs. Dans un premier temps, nous avons étudié les conditions de culture pour l isolement et l expansion des CSMs, et nous avons choisi les conditions de culture optimales pour ces CSMs. Dans un deuxième temps, nous avons étudié l effet du TGF-ß1 sur les CSMs et les chondrocytes. Nous avons trouvé que la phosphorylation de Smad3 est augmentée après 1 heur de stimulation avec du TGF-ß1, mais diminuée après 3 heures: Au même temps, Smad3p s est translocalisée dans le noyau. C est un effet dépendant de la concentration. L expression du gène de Sox9 est augmentée également après stimulation avec du TGF-ß1. Dans un troisième temps, nous avons étudié l effet de la contrainte d étirement sur les CSMs et les chondrocytes. Nous avons trouvé que la contrainte d étirement avait le même effet que le TGF-ß1 sur la phosphorylation de Smad3 et l expression du gène de Sox9. En plus, avec l étirement, le TGF-ß1 latent est activé, le RII est internalisé et les CSMs sont plus alongées. De plus, il y avait un effet synergique entre l étirement et le TGF-ß1. En conclusion, la contrainte d étirement active la différenciation des CSMs par l activation de la voie du TGF-ß. Il semble que le mécanisme de mécanotransduction est contrôlé par plusieurs facteurs.Transforming growth factor-ß (TGF-ß) is a key factor for chondrogenic differentiation of human mesenchymal stem cells (hMSCs). Mechanical loading has also been shown to be important in MSC differentiation, but mechanisms by which mechanotransduction occurs remain largely elusive. The aim of our work was to define the short-time effect of cyclic tensile strain (CTS, 0.5 Hz or 1 Hz, 5% equibiaxial strain) on the hMSCs. Firstly, we investigated the culture parameters for the isolation of hMSCs and by which the initial expansion was influenced. We chose the opitimal culture conditions for hMSCs. Secondly, we studied the effect of TGF-ß1 on the hMSCs and chondrocytes. We found that Smad3 phosphorylation (Smad3p) was increased after 1 hour by the stimulation of TGF-ß1, but decreased after 3 hours, Smad3p translocation to the nucleus was observed too, and the effect was dose-dependent. At the same time, TGF-ß1 increased the expression of the gene of Sox9. Finally, we studied the effect of CTS on the hMSCs and chondrocytes. Results showed that CTS had the same effect that TGF-ß1 on the Smad3 phosphorylation and the expression of the gene of Sox9: CTS increases the latent TGF-ß1 in the medium, increases the internalisation of the type II TGF-ß receptor and changes the form of hMSCs. Moreover a synergy effect between CTS and TGF-ß1 was observed. We conclude that CTS could stimulate MSC differentiation by the activation of endogenous TGF-ß signaling pathway. It seems that the mechanisms of mechanotransduction are controlled by multiple factors.NANCY1-Bib. numérique (543959902) / SudocSudocFranceF

Pre-professional international mobility of European pharmacy students - a French example.

Internationalisation, as well as the need to interact with international partners in academia and in the pharmaceutical industry, brings an international experience to the pharmacist's career, which is essential. The objective of present work is to provide a preliminary study of the current situation of the pre-professional mobility of pharmacy students. It represents the first case study of the international pre-professional mobility of pharmacy students in France, and in north-eastern France in particular. The study is based on a recent preliminary survey among pharmacy students, conducted in 2020 at the University of Lorraine's Faculty of Pharmacy, reflecting the impact of international mobility programmes, such as the European Union educational and training mobility programme Erasmus+, on the pharmacy curriculum. The results of the present work tend to show that, despite a number of barriers to the international mobility of pharmacy students, the outcomes of international pre-professional mobility are rather positive in their globality

Analyses of the effects of collection and processing time on the results of serology testing of cadaveric cornea donors

International audienc

Systematic Review on CAR-T Cell Clinical Trials Up to 2022: Academic Center Input

The development of Chimeric Antigen Receptor T cells therapy initiated by the United States and China is still currently led by these two countries with a high number of clinical trials, with Europe lagging in launching its first trials. In this systematic review, we wanted to establish an overview of the production of CAR-T cells in clinical trials around the world, and to understand the causes of this delay in Europe. We particularly focused on the academic centers that are at the heart of research and development of this therapy. We counted 1087 CAR-T cells clinical trials on ClinicalTrials.gov (Research registry ID: reviewregistry1542) on the date of 25 January 2023. We performed a global analysis, before analyzing the 58 European trials, 34 of which sponsored by academic centers. Collaboration between an academic and an industrial player seems to be necessary for the successful development and application for marketing authorization of a CAR-T cell, and this collaboration is still cruelly lacking in European trials, unlike in the leading countries. Europe, still far behind the two leading countries, is trying to establish measures to lighten the regulations surrounding ATMPs and to encourage, through the addition of fundings, clinical trials involving these treatments

Systematic Review on CAR-T Cell Clinical Trials Up to 2022: Academic Center Input

The development of Chimeric Antigen Receptor T cells therapy initiated by the United States and China is still currently led by these two countries with a high number of clinical trials, with Europe lagging in launching its first trials. In this systematic review, we wanted to establish an overview of the production of CAR-T cells in clinical trials around the world, and to understand the causes of this delay in Europe. We particularly focused on the academic centers that are at the heart of research and development of this therapy. We counted 1087 CAR-T cells clinical trials on ClinicalTrials.gov (Research registry ID: reviewregistry1542) on the date of 25 January 2023. We performed a global analysis, before analyzing the 58 European trials, 34 of which sponsored by academic centers. Collaboration between an academic and an industrial player seems to be necessary for the successful development and application for marketing authorization of a CAR-T cell, and this collaboration is still cruelly lacking in European trials, unlike in the leading countries. Europe, still far behind the two leading countries, is trying to establish measures to lighten the regulations surrounding ATMPs and to encourage, through the addition of fundings, clinical trials involving these treatments