The successful application of mesenchymal stem cells (MSCs) remains a major challenge in stem cell therapy. Currently, several in vitro studies have indicated potentially beneficial interactions of MSCs with immunosuppressive drugs. These interactions can be even more complex in vivo, and it is in this setting that we investigate the effect of MSCs in combination with Cyclosporine A (CsA) on transplantation reaction and allogeneic cell survival. Using an in vivo mouse model, we found that CsA significantly promoted the survival of MSCs in various organs and tissues of the recipients. In addition, compared to treatment with CsA or MSCs alone, the survival of transplanted allogeneic cells was significantly improved after the combined application of MSCs with CsA. We further observed that the combinatory treatment suppressed immune response to the alloantigen challenge and modulated the immune balance by harnessing proinflammatory CD4+T-bet+ and CD4+RORγt+ cell subsets. These changes were accompanied by a significant decrease in IL-17 production along with an elevated level of IL-10. Co-cultivation of purified naive CD4+ cells with peritoneal macrophages isolated from mice treated with MSCs and CsA revealed that MSC-educated macrophages play an important role in the immunomodulatory effect observed on distinct T-cell subpopulations. Taken together, our findings suggest that CsA promotes MSC survival in vivo and that the therapeutic efficacy of the combination of MSCs with CsA is superior to each monotherapy. This combinatory treatment thus represents a promising approach to reducing immunosuppressant dosage while maintaining or even improving the outcome of therapy.
- MeSH
- alografty účinky léků imunologie MeSH
- cyklosporin farmakologie terapeutické užití MeSH
- cytokiny metabolismus MeSH
- imunosupresiva farmakologie terapeutické užití MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- preklinické hodnocení léčiv MeSH
- přežívání štěpu účinky léků imunologie MeSH
- T-lymfocyty účinky léků metabolismus MeSH
- transplantace mezenchymálních kmenových buněk * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The immunosuppressive effects of systemically administered mesenchymal stem cells (MSCs) and immunosuppressive drugs have been well documented. We analysed the mechanisms underlying the therapeutic effect of MSCs applied locally in combination with non-specific immunosuppression in a mouse model of allogeneic skin transplantation. The MSC-seeded and cyclosporine A (CsA)-loaded nanofibre scaffolds were applied topically to skin allografts in a mouse model and the local immune response was assessed and characterized. MSCs migrated from the scaffold into the side of injury and were detected in the graft region and draining lymph nodes (DLNs). The numbers of graft-infiltrating macrophages and the production of nitric oxide (NO) were significantly decreased in recipients treated with MSCs and CsA, and this reduction correlated with impaired production of IFNγ in the graft and DLNs. In contrast, the proportion of alternatively activated macrophages (F4/80+CD206+cells) and the production of IL-10 by intragraft macrophages were significantly upregulated. The ability of MSCs to alter the phenotype of macrophages from the M1 type into an M2 population was confirmed in a co-culture system in vitro. We suggest that the topical application of MSCs in combination with CsA induces a switch in macrophages to a population with an alternatively activated 'healing' phenotype and producing elevated levels of IL-10. These alterations in macrophage phenotype and function could represent one of the mechanisms of immunosuppressive action of MSCs applied in combination with CsA. Copyright © 2015 John Wiley & Sons, Ltd.
- MeSH
- alografty MeSH
- buněčná diferenciace MeSH
- cyklosporin farmakologie MeSH
- makrofágy metabolismus MeSH
- mezenchymální kmenové buňky metabolismus MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- přežívání štěpu účinky léků MeSH
- transplantace kůže * MeSH
- transplantace mezenchymálních kmenových buněk * MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Immunosuppressive drugs are widely used to treat undesirable immune reaction, however their clinical use is often limited by harmful side effects. The combined application of immunosuppressive agents with mesenchymal stem cells (MSCs) offers a promising alternative approach that enables the reduction of immunosuppressive agent doses and simultaneously maintains or improves the outcome of therapy. The present study aimed to determinate the effects of immunosuppressants on individual T cell subpopulations and to investigate the efficacy of MSC-based treatment combined with immunosuppressive drugs. We tested the effect of five widely used immunosuppressants with different action mechanisms: cyclosporine A, mycophenolate mofetil, rapamycin, and two glucocorticoids - prednisone and dexamethasone in combination with MSCs on mouse CD4+ and CD8+ lymphocyte viability and activation, Th17 (RORγt+), Th1 (T-bet+), Th2 (GATA-3+) and Treg (Foxp3+) cell proportion and on the production of corresponding key cytokines (IL-17, IFNγ, IL-4 and IL-10). We showed that MSCs modulate the actions of immunosuppressants and in combination with immunosuppressive drugs display distinct effect on cell activation and balance among different T lymphocytes subpopulations and exert a suppressive effect on proinflammatory T cell subsets while promoting the functions of anti-inflammatory Treg lymphocytes. The results indicated that MSC-based therapy could be a powerful strategy to attenuate the negative effects of immunosuppressive drugs on the immune system.
- MeSH
- aktivace lymfocytů účinky léků imunologie MeSH
- cyklosporin farmakologie MeSH
- cytokiny metabolismus MeSH
- dexamethason farmakologie MeSH
- glukokortikoidy farmakologie MeSH
- imunosupresiva farmakologie MeSH
- kokultivační techniky MeSH
- kultivované buňky MeSH
- kyselina mykofenolová farmakologie MeSH
- mezenchymální kmenové buňky cytologie imunologie metabolismus MeSH
- myši inbrední BALB C MeSH
- prednison farmakologie MeSH
- proliferace buněk účinky léků MeSH
- průtoková cytometrie MeSH
- sirolimus farmakologie MeSH
- T-lymfocyty - podskupiny cytologie účinky léků imunologie MeSH
- transplantace mezenchymálních kmenových buněk metody MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- hemoragická horečka Ebola prevence a kontrola MeSH
- hygiena práce * MeSH
- kontrola infekčních nemocí metody MeSH
- lidé MeSH
- ochranné oděvy využití MeSH
- ochranné prostředky očí normy využití MeSH
- ochranné rukavice využití MeSH
- přenos infekce z pacienta na zdravotnického pracovníka prevence a kontrola MeSH
- prostředky na ochranu dýchání klasifikace normy využití MeSH
- zdravotnický personál * zákonodárství a právo MeSH
- Check Tag
- lidé MeSH
