UNLABELLED: Stem cell-based therapy has become an attractive and promising approach for the treatment of severe injuries or thus-far incurable diseases. However, the use of stem cells is often limited by a shortage of available tissue-specific stem cells; therefore, other sources of stem cells are being investigated and tested. In this respect, mesenchymal stromal/stem cells (MSCs) have proven to be a promising stem cell type. In the present study, we prepared MSCs from bone marrow (BM-MSCs) or adipose tissue (Ad-MSCs) as well as limbal epithelial stem cells (LSCs), and their growth, differentiation, and secretory properties were compared. The cells were grown on nanofiber scaffolds and transferred onto the alkali-injured eye in a rabbit model, and their therapeutic potential was characterized. We found that BM-MSCs and tissue-specific LSCs had similar therapeutic effects. Clinical characterization of the healing process, as well as the evaluation of corneal thickness, re-epithelialization, neovascularization, and the suppression of a local inflammatory reaction, were comparable in the BM-MSC- and LSC-treated eyes, but results were significantly better than in injured, untreated eyes or in eyes treated with a nanofiber scaffold alone or with a nanofiber scaffold seeded with Ad-MSCs. Taken together, the results show that BM-MSCs' therapeutic effect on healing of injured corneal surface is comparable to that of tissue-specific LSCs. We suggest that BM-MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain. SIGNIFICANCE: Damage of ocular surface represents one of the most common causes of impaired vision or even blindness. Cell therapy, based on transplantation of stem cells, is an optimal treatment. However, if limbal stem cells (LSCs) are not available, other sources of stem cells are tested. Mesenchymal stem cells (MSCs) are a convenient type of cell for stem cell therapy. The therapeutic potential of LSCs and MSCs was compared in an experimental model of corneal injury, and healing was observed following chemical injury. MSCs and tissue-specific LSCs had similar therapeutic effects. The results suggest that bone marrow-derived MSCs can be used for ocular surface regeneration in cases when autologous LSCs are absent or difficult to obtain.

• MeSH
• biologické markery metabolismus   MeSH
• buněčná a tkáňová terapie metody   MeSH
• buněčná diferenciace MeSH
• buňky kostní dřeně cytologie   fyziologie   MeSH
• chemické popálení patologie   terapie   MeSH
• epitelové buňky cytologie   fyziologie   transplantace   MeSH
• exprese genu MeSH
• fyziologická neovaskularizace MeSH
• králíci MeSH
• limbus corneae krevní zásobení   zranění   MeSH
• mezenchymální kmenové buňky cytologie   fyziologie   MeSH
• primární buněčná kultura MeSH
• proliferace buněk MeSH
• reepitalizace fyziologie   MeSH
• rohovkový epitel krevní zásobení   zranění   MeSH
• tkáňové podpůrné struktury MeSH
• transplantace mezenchymálních kmenových buněk * MeSH
• tuková tkáň cytologie   fyziologie   MeSH
• tukové buňky cytologie   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

B cells play an important role in the immune responses which affect the outcomes of kidney allografts. Dynamic changes of B-cell compartments in clinical kidney transplantation are still poorly understood. B-cell subsets were prospectively monitored using flow cytometry for 1 year in 98 kidney transplant recipients. Data were correlated with immunosuppression and clinical outcomes. An increase in the total population of B lymphocytes was observed during the first week after transplantation. The level of IgM(high) CD38(high) CD24(high) transitional B cells reduced significantly up until the third month, with partial repopulation in the first year. Lower numbers of transitional B cells in the third month were associated with higher risk of graft rejection. IgM(+) IgD(+) CD27(-) naive B cells did not change within follow-up. IgM(+) CD27(+) nonswitched memory B cells and IgM(-) CD27(+) switched memory B cells increased on post-operative day 7. IgM(-) CD38(high) CD27(high) plasmablasts showed similar kinetics during the first post-transplant year, similar to transitional B cells. In conclusion, sensitized kidney transplant recipients as well as those with either acute or chronic rejection within the first post-transplant year exhibited lower levels of transitional B cells. Therefore, these data further support the hypothesis that transitional B cells have a protective role in kidney transplantation.

• MeSH
• alografty MeSH
• antigen CD24 metabolismus   MeSH
• antigeny CD27 metabolismus   MeSH
• antigeny CD38 metabolismus   MeSH
• časové faktory MeSH
• dítě MeSH
• dospělí MeSH
• imunologická paměť imunologie   MeSH
• imunosupresivní léčba MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• plazmatické buňky imunologie   MeSH
• podskupiny B-lymfocytů imunologie   MeSH
• předškolní dítě MeSH
• prekurzorové B-lymfoidní buňky imunologie   MeSH
• příjemce transplantátu MeSH
• prospektivní studie MeSH
• rejekce štěpu imunologie   MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• transplantace ledvin * MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The increased demand for kidney transplantation and organ shortage resulted in the increased use of kidneys from suboptimal donors. Therefore, identification of kidneys that can be accepted without significantly compromising the outcome of allograft or recipient has become critical. A robust assessment of organ quality is of particular importance especially in kidneys from elderly donors in whom morphological and functional changes associated with aging and diseases are obvious. A number of predictive tools have been developed to help with evaluating the suitability of a deceased-donor kidney for transplantation. Among those, Kidney Donor Profile Index and zero hour graft biopsy in elderly donors have been already implemented in several transplant programs. This review captures the recent literature on this subject and discusses approaches for evaluating the quality of kidney grafts from elderly donors.

• MeSH
• biologické markery analýza   MeSH
• biopsie MeSH
• dárci tkání * MeSH
• hodnocení rizik * MeSH
• lidé středního věku MeSH
• lidé MeSH
• přežívání štěpu MeSH
• rejekce štěpu MeSH
• senioři MeSH
• seznamy čekatelů MeSH
• transplantace ledvin * MeSH
• výběr dárců * MeSH
• výběr pacientů MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

V tomto projektu budeme pomocí molekulárně genetických a imunologických metod zkoumat roli fenotypů B lymfocytů a jejich transkriptů v periferní krvi a dárcovsky specifických protilátek při alloimunitní odpovědi v kohortě nemocných po transplantaci ledviny a v experimentálních modelech. Předpokládáme, že specifické populace B lymfocytů a jejich transkripty budou asociovány s absencí rejekce, kdežto jiné s odhojením štěpů. Projekt tak může pomoci identifikovat nové biomarkery použitelné pro budoucí validaci v klinických studiích s cílem individualizovat imunosupresivní terapii.; In this project, we will investigate the role of B-lymphocytes phentotypes and their transcripts in peripheral blood and the donor specific antibodies in alloimune response in the cohort of kidney transplant recipients, and in experimental models by means of molecular genetic and immunological methods. We assume that the specific populations of B-lymphocytes and their transcripts will be associated with the absence of rejection, whereas others with the rejection of grafts. The project can so help to identify new biomarkers applicable for the future validation in clinical studies with the objective to individualize the immunosuppressive therapy.

• MeSH
• B-lymfocyty MeSH
• transplantace ledvin MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• nefrologie
• alergologie a imunologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

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

The immunoregulatory properties of mesenchymal stem cells (MSCs) have been well documented in various models in vitro and in vivo. Furthermore, a population of regulatory B cells (Bregs) that produce relatively high concentrations of IL-10 has been recently described. To study the relationship between MSCs and Bregs, we analyzed the effects of MSCs on IL-10 production by lipopolysaccharide (LPS)-activated mouse B cells. The production of IL-10 by B cells remained preserved in the presence of MSCs and was even significantly enhanced by IFN-γ. However, the production of IL-10 was strongly suppressed in cultures containing MSCs and IFN-γ. Preincubation of MSCs, but not of B cells, with IFN-γ induced the suppression of IL-10 secretion in cultures containing MSCs and B cells. The supernatants from IFN-γ-treated MSCs had no inhibitory effect, and the suppression of IL-10 production was abrogated if the MSCs and B cells were separated in a transwell system. Analysis of the gene expression of IFN-γ- or IFN-γ and LPS-treated MSCs revealed a strong upregulation of genes for indoleamine-2,3-dioxygenase (IDO), cyclooxygenase-2 (Cox-2) and programmed cell death-ligand 1 (PD-L1). While the inhibition of IDO activity or the inclusion of the neutralization monoclonal antibody anti-PD-L1 did not abrogate the suppression, indomethacin, an inhibitor of Cox-2, completely inhibited the MSC-mediated suppression of IL-10 production. Accordingly, the production of IL-10 by B cells was inhibited by exogenous prostaglandin E2. The results thus suggest that IFN-γ-treated MSCs strongly inhibit IL-10 production by activated B cells by a mechanism requiring cell contact and involving the Cox-2 pathway.

• MeSH
• aktivace lymfocytů účinky léků   MeSH
• antigeny CD274 antagonisté a inhibitory   genetika   imunologie   MeSH
• B-lymfocyty cytologie   účinky léků   imunologie   MeSH
• cyklooxygenasa 2 genetika   imunologie   MeSH
• difuzní komory kultivační MeSH
• dinoproston farmakologie   MeSH
• indolamin-2,3,-dioxygenasa genetika   imunologie   MeSH
• indomethacin farmakologie   MeSH
• inhibitory cyklooxygenasy farmakologie   MeSH
• interferon gama farmakologie   MeSH
• interleukin-10 antagonisté a inhibitory   genetika   imunologie   MeSH
• kokultivační techniky MeSH
• kultivační média speciální farmakologie   MeSH
• lipopolysacharidy farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   účinky léků   imunologie   MeSH
• mezibuněčná komunikace imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• neutralizující protilátky farmakologie   MeSH
• primární buněčná kultura MeSH
• regulace genové exprese MeSH
• signální transdukce MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Induction therapy can improve kidney transplantation (KTx) outcomes, but little is known about the mechanisms underlying its effects. METHODS: The mRNA levels of T cell-related genes associated with tolerance or rejection (CD247, GZMB, PRF1, FOXP3, MAN1A1, TCAIM, and TLR5) and lymphocyte subpopulations were monitored prospectively in the peripheral blood of 60 kidney transplant recipients before and 7, 14, 21, 28, 60, 90 days, 6 months, and 12 months after KTx. Patients were treated with calcineurin inhibitor-based triple immunosuppression and induction with rabbit anti-thymocyte globulin (rATG, n = 24), basiliximab (n = 17), or without induction (no-induction, n = 19). A generalized linear mixed model with gamma distribution for repeated measures, adjusted for rejection, recipient/donor age and delayed graft function, was used for statistical analysis. RESULTS: rATG treatment caused an intense reduction in all T cell type population and natural killer (NK) cells within 7 days, then a slow increase and repopulation was observed. This was also noticed in the expression levels of CD247, FOXP3, GZMB, and PRF1. The basiliximab group exhibited higher CD247, GZMB, FOXP3 and TCAIM mRNA levels and regulatory T cell (Treg) counts than the no-induction group. The levels of MAN1A1 and TLR5 mRNA expressions were increased, whereas TCAIM decreased in the rATG group as compared with those in the no-induction group. CONCLUSION: The rATG induction therapy was associated with decreased T and NK cell-related transcript levels and with upregulation of two rejection-associated transcripts (MAN1A1 and TLR5) shortly after KTx. Basiliximab treatment was associated with increased absolute number of Treg cells, and increased level of FOXP3 and TCAIM expression.

• MeSH
• antigeny CD3 genetika   MeSH
• antilymfocytární sérum terapeutické užití   MeSH
• dospělí MeSH
• exprese genu účinky léků   MeSH
• forkhead transkripční faktory genetika   MeSH
• granzymy genetika   MeSH
• imunologická tolerance účinky léků   genetika   MeSH
• imunosupresiva terapeutické užití   MeSH
• imunosupresivní léčba metody   MeSH
• indukční chemoterapie metody   MeSH
• inhibitory kalcineurinu terapeutické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mannosidasy genetika   MeSH
• messenger RNA krev   MeSH
• mladý dospělý MeSH
• monoklonální protilátky terapeutické užití   MeSH
• NKT buňky účinky léků   imunologie   MeSH
• perforin genetika   MeSH
• počet lymfocytů MeSH
• prospektivní studie MeSH
• regulační T-lymfocyty účinky léků   imunologie   MeSH
• rejekce štěpu genetika   imunologie   prevence a kontrola   MeSH
• rekombinantní fúzní proteiny terapeutické užití   MeSH
• senioři MeSH
• transplantace ledvin metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this study was to examine whether nanofiber scaffolds seeded with rabbit bone marrow mesenchymal stem cells (MSCs nanofibers) transferred onto the damaged corneal surface and covered with cyclosporine A (CsA)-loaded nanofiber scaffolds (CsA nanofibers) enable healing of the rabbit cornea injured with 1N NaOH. The healing of damaged corneas was examined morphologically, immunohistochemically and biochemically on day 24 after the injury. Compared to untreated injured corneas, where corneal ulceration or large corneal thinning or even perforation were developed, injured corneas treated with drug free nanofibers healed without profound disturbances in a majority of cases, although with fibrosis and scar formation. In injured corneas treated with CsA nanofibers or MSCs nanofibers, the development of scar formation was reduced. Best healing results were obtained with a combination of MSCs and CsA nanofibers (MSCs-CsA nanofibers). Corneas healed with highly restored transparency. Neovascularization highly expressed in untreated injured corneas and reduced in corneas treated with CsA nanofibers or MSCs nanofibers, was suppressed in corneas treated with MSCs-CsA nanofibers. The levels of matrix metalloproteinase 9, inducible nitric oxide synthase, interleukin 6, α-smooth muscle actin, tumor growth factor β and vascular endothelial growth factor were significantly decreased in these corneas as compared to untreated corneas, where the levels of the above mentioned markers were high. In conclusion, MSCs-CsA nanofibers were effective in the treatment of severe alkali-induced corneal injury.

• MeSH
• cyklosporin aplikace a dávkování   MeSH
• hojení ran účinky léků   MeSH
• hydroxid sodný toxicita   MeSH
• imunosupresiva aplikace a dávkování   MeSH
• jizva prevence a kontrola   MeSH
• kaustika toxicita   MeSH
• králíci MeSH
• lékové transportní systémy metody   MeSH
• modely nemocí na zvířatech MeSH
• nanovlákna MeSH
• neovaskularizace rohovky prevence a kontrola   MeSH
• nosiče léků MeSH
• poranění rohovky terapie   MeSH
• tkáňové podpůrné struktury MeSH
• transplantace mezenchymálních kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• Publikační typ
• abstrakt z konference MeSH

The significance of borderline changes in kidney allograft biopsies is widely debated. To help resolve this, we studied differences in intrarenal gene expression patterns between early clinical and 3-month protocol biopsies, all of which had borderline histologic changes. The gene expression profiles in training set of patients by microarray analysis and data were validated in a larger cohort using RT-qPCR. There was greater expression of immunity- and inflammation-related genes in the early clinical biopsies compared to the 3-month protocol biopsies with borderline changes. In early clinically manifested borderline changes, graft deterioration within 24 months due to chronic rejection was associated with increased activation of immune, defense, and inflammatory processes. Regression modeling identified higher donor age and expression of macrophage receptor CLEC5A as risk factors for progression. In the 3-month protocol biopsies with borderline changes, graft dysfunction was associated with increased expression of fibrinogen complex transcripts. The discrimination power of fibrinogen was confirmed by cross-validation on two independent cohorts. Thus, our study highlights variations in gene expression between clinical and subclinical borderline changes despite similar histological findings. The data also support a recommendation for frequent patient monitoring, especially in those with borderline changes who received grafts from older donors.

• MeSH
• asymptomatické nemoci MeSH
• biopsie MeSH
• časná diagnóza MeSH
• časové faktory MeSH
• diagnostické techniky molekulární * MeSH
• dospělí MeSH
• fenotyp MeSH
• genetická predispozice k nemoci MeSH
• genetické markery * MeSH
• hodnocení rizik MeSH
• ledviny patologie   patofyziologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• polymerázová řetězová reakce MeSH
• prediktivní hodnota testů MeSH
• regulace genové exprese MeSH
• rejekce štěpu genetika   patologie   patofyziologie   MeSH
• reprodukovatelnost výsledků MeSH
• retrospektivní studie MeSH
• rizikové faktory MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• senioři MeSH
• stanovení celkové genové exprese MeSH
• transplantace ledvin škodlivé účinky   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• pozorovací studie MeSH
• práce podpořená grantem MeSH
• validační studie MeSH