Rapamycin prevents experimental sclerodermatous chronic graft-versus-host disease in mice

Background: The most widely used mice model of chronic graft-versus-host disease (cGvHD) is an MHC-matched bone marrow transplantation model of sclerodermatous cGvHD. A limitation of that model is that mortality is relatively low, making difficult to study the impact of potentially therapeutic compounds. Aims: To develop a more severe model of cGVHD and to assess the impact of Rapamycin administration in that model. Results: Lethally irradiated Balb/C mice were injected with 10x106 bone marrow cells and 70x106 splenocytes from B10.D2 donor mice. Twenty-one days later, all mice developed cGvHD. For the severe model, donor B10.D2 mice were injected with 0.5x106 splenocytes from Balb/C twenty-one days before transplantation. All mice from the severe model (n=8) died a median of 32 days while 3 of 7 mice in the classical model survived beyond day 52. Mean survival was decreased in the severe model compared to the classical model (32 days versus 37 days; p=0.0185). Recipient mice in the severe group experienced higher weight loss, hair loss and skin fi brosis. Numbers of T lymphocytes (231.9 ± 151.4 versus 951 ± 532.8; p=0.0032) and CD4+ T cells (63.25 ± 41.93 versus 135.0 ± 14.39; p=0.0018) per microliter of blood at day 21 were lower in the severe group than in the classical model. Moreover, number of regulatory T cells (Tregs) was decreased in the severe model (1.250 ± 0.8864 versus 8.000 ± 6.753; p=0.0151). We then investigated whether rapamycin administration could prevent GVHD in the severe model. All (n=8) mice treated with PBS (placebo) died a median of 32 days after transplantation, while 6 of 8 mice given 1 mg/kg/day i.p. rapamycin survived beyond day 52 (p=0.0012). Number of Tregs/μl was higher at day 21 in rapamycin-treated mice than in mice given PBS (2.000±1.195 versus 1.250±0.8864; p=0.0796). Moreover, number of naïve CD4+T (10.00±4.192 versus 30.25±5.185; p= 0.0089) and effector memory T cells (EMT) (30.67±3.180 versus 67.33±7.881; p= 0.0125) were higher in rapamycin mice. Finally, proliferation of EMT (assessed by fl ow cytometry using Ki-67) was higher in PBS than in rapamycin mice (45.28%±4.084 versus 31.90%± 2.003; p=0.0474). Conclusion: We have developed a mice model of severe cGVHD. Interestingly, rapamycin prevented death from cGVHD in that model, perhaps through in vivo expansion of Treg

Reconstitution of adaptive immunity after umbilical cord blood transplantation and clinical implication regarding risk of infections

In comparison with allogeneic stem cell transplantation (alloHSCT) with other stem cell sources, umbilical cord blood transplantation (UCBT) was traditionally associated with increased risk of infections, particularly during the first 3 months after transplantation. Longitudinal studies of immune monitoring reported peculiar patterns of T- and B-cell recovery in the peripheral blood of UCB recipients during the first months post-transplantation. Overall, current data suggest delayed reconstitution of naive and memory CD4+ and CD8+ T-cell pools after UCBT. This is particularly true for adult recipients and for patients who received in vivo T-cell depleting approaches before the transplantation. Such delayed T-cell recovery may increase susceptibility of UCB recipients for developing opportunistic infections and viral reactivations. Regarding B-cell recovery, UCBT was associated with accelerated B-lymphopoiesis. Recent studies also reported evidence for faster functional memory B-cell recovery in UCB recipients. In this article, we briefly review T- and B-cell reconstitution after alloHSCT, with emphasis on peculiarities observed after UCBT. We further put these data in lines with risks of infections after UCBT

What Is The Role For Regulatory T-Cells After Nonmyeloablative Conditioning?

peer reviewe

Comparison of Mesenchymal Stromal Cells From Different Origins for the Treatment of Graft-vs.-Host-Disease in a Humanized Mouse Model

Mesenchymal stromal cells (MSCs) have potent immunomodulatory properties that make them an attractive tool against graft- vs.-host disease (GVHD). However, despite promising results in phase I/II studies, bone marrow (BM-) derived MSCs failed to demonstrate their superiority over placebo in the sole phase III trial reported thus far. MSCs from different tissue origins display different characteristics, but their therapeutic benefits have never been directly compared in GVHD. Here, we compared the impact of BM-, umbilical cord (UC-), and adipose-tissue (AT-) derived MSCs on T-cell function in vitro and assessed their efficacy for the treatment of GVHD induced by injection of human peripheral blood mononuclear cells in NOD-scid IL-2Rγnull HLA-A2/HHD mice. In vitro, resting BM- and AT-MSCs were more potent than UC-MSCs to inhibit lymphocyte proliferation, whereas UC- and AT-MSCs induced a higher regulatory T-cell (CD4+CD25+FoxP3+)/T helper 17 ratio. Interestingly, AT-MSCs and UC-MSCs activated the coagulation pathway at a higher level than BM-MSCs. In vivo, AT-MSC infusions were complicated by sudden death in 4 of 16 animals, precluding an analysis of their efficacy. Intravenous MSC infusions (UC- or BM- combined) failed to significantly increase overall survival (OS) in an analysis combining data from 80 mice (hazard ratio [HR] = 0.59, 95% confidence interval [CI] 0.32–1.08, P = 0.087). In a sensitivity analysis we also compared OS in control vs. each MSC group separately. The results for the BM-MSC vs. control comparison was HR = 0.63 (95% CI 0.30–1.34, P = 0.24) while the figures for the UC-MSC vs. control comparison was HR = 0.56 (95% CI 0.28–1.10, P = 0.09). Altogether, these results suggest that MSCs from various origins have different effects on immune cells in vitro and in vivo. However, none significantly prevented death from GVHD. Finally, our data suggest that the safety profile of AT-MSC and UC-MSC need to be closely monitored given their pro-coagulant activities in vitro

Study of imune reconstitution after hematopoetic stem cell transplantation : focus on thymic function and regulatory T cells

L’allogre e de cellules souches hématopoïétiques, après conditionnement myéloalbatif ou non, est devenue une option de choix dans le traitement des maladies hématologiques malignes ou génétiques. Malheureusement le succès de ce traitement se trouve trop souvent entaché par des complications majeures telles que les infections ou la maladie du greffon contre l’hôte (GVHD), dans laquelle les cellules du donneur attaquent l’organisme du receveur. La reconstitution d’un système immunitaire fonctionnel, notamment via le thymus, est une étape clef dans la résolution de ces deux complications et l’utilisation de lymphocytes T régulateurs semble très prometteuse dans la lutte contre la GVHD. Le présent travail vise donc à étudier la reconstitution du système immunitaire après allogreffe sous trois angles différents, en nous focalisant sur la fonction thymique et l’influence des lymphocytes T régulateurs (Treg). Dans la première partie, nous évaluons la reconstitution immunitaire à long terme après greffe non-myéloablative et montrons que la néo-génération de lymphocytes T par le thymus s’effectue à partir du jour 100 chez les patients de moins de 60 ans alors qu’au delà de cet âge, elle est quasiment absente. Nous établissons également que la GVHD chronique (cGVHD) sévère possède des effets extrêmement délétères sur le thymus. Étant donné l’impact de la cGVHD sur le thymus et la reconstitution immunitaire, nous avons voulu comparer l’impact, sur ces deux paramètres, d’un conditionnement nonmyéloalbatif censé réduire la GVHD et composé d’une irradiation lymphoïde totale et d’anticorps anti-thymocytes (TLI-ATG) par rapport au conditionnement classique consistant en une irradiation corporelle totale associée à de la fludarabine (TBI-Flu). Il ressort de cette étude que, bien que réduisant de manière significative l’incidence de cGVHD, le conditionnement TLI-ATG altère fortement la reconstitution du système immunitaire et impacte négativement le taux de rechute/progression chez les patients traités. Enfin, toujours dans le but de réduire le taux de GVHD post-greffe, nous avons choisi d’examiner l’impact des Treg comme traitement de la xGVHD. Dans ce but, nous montrons qu’une sélection de Treg humains, sur CliniMACS par déplétion CD8/19 et sélection CD25, fournit une population de cellules enrichies en Treg capable de retarder de manière statistiquement significative l’apparition d’une GVHD xénogénique induite par injection de PBMC humains (autologues aux Treg) dans un modèle pré-clinique chez la souris NSG. De plus, lorsque injectée sans les PBMC, cette population enrichie n’induit pas par elle-même de GHVD.Etude de la reconstutition immunitaire après greffes de cellules souches hématopoïétique

Genetics of severe early-onset obesity: diagnosis of monogenic etiologies as a new paradigm and challenge in the era of new anti-obesity therapies

editorial reviewedRare genetic forms of obesity are mainly caused by impaired energy homeostasis with dysregulation in eating behavior and energy expenditure involving hypothalamic pathways. Recent advances showed that more than 60 genes coding for proteins located in the hypothalamic leptin/melanocortin pathway contribute to the development of these rare forms of obesity, syndromic and monogenic. A summary of these new insights in monogenic obesity is presented here to help physicians in the understanding on the new issues of the diagnosis of genetic obesity. The availability of new anti-obesity therapies opens the field for current and future perspectives of personalized medicine approach