The retina represents a highly specialized structure with the primary function to capture a light signal and to convert it into electrical impulses. Any damage or disease of the retina can cause visual impairment. Since retinal degenerative diseases are generally associated with immune cell infiltration, a local inflammatory reaction, and cytokine burn, there is a need for mechanisms to prevent the retina from damage by a deleterious immune reaction. In this study, we show that mouse retinal explants co-cultivated with stimulated spleen cells, inhibit in a dose-dependent manner the activation of T cells, and suppress the production of cytokines interleukin-2, interleukin-10, and interferon-[Formula: see text]. The immunoregulatory properties of the retina were mainly mediated by a paracrine effect since retinal explants, separated by a semipermeable membrane, or supernatants obtained after the cultivation of retinal explants, inhibited the reactivity of immune cells. A model of retinal damage was established by the application of sodium iodate which selectively destroys photoreceptors, as it was demonstrated by a decrease in the number of rhodopsin-positive cells. This process was accompanied by increased infiltration of the retina with cells of the immune system and by a local inflammatory reaction. The pharmacologically damaged retina had significantly decreased the ability to inhibit T cell activation and production of cytokines by immune cells. Overall, the results showed that the retina possesses immunoregulatory properties and inhibits the activation and functions of T cells. However, the immunomodulatory properties of the retina are decreased if the retina is damaged.

• Klíčová slova
• cytokines, healthy retina, immunosuppression, inflammatory eye, pharmacologically damaged retina,
• MeSH
• cytokiny * metabolismus   MeSH
• myši MeSH
• retina * MeSH
• zánět metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytokiny * MeSH

Negative impacts of nanomaterials on stem cells and cells of the immune system are one of the main causes of an impaired or slowed tissue healing. Therefore, we tested effects of four selected types of metal nanoparticles (NPs): zinc oxide (ZnO), copper oxide (CuO), silver (Ag), and titanium dioxide (TiO2) on the metabolic activity and secretory potential of mouse mesenchymal stem cells (MSCs), and on the ability of MSCs to stimulate production of cytokines and growth factors by macrophages. Individual types of nanoparticles differed in the ability to inhibit metabolic activity, and significantly decreased the production of cytokines and growth factors (interleukin-6, vascular endothelial growth factor, hepatocyte growth factor, insulin-like growth factor-1) by MSCs, with the strongest inhibitory effect of CuO NPs and the least effect of TiO2 NPs. The recent studies indicate that immunomodulatory and therapeutic effects of transplanted MSCs are mediated by macrophages engulfing apoptotic MSCs. We co-cultivated macrophages with heat-inactivated MSCs which were untreated or were preincubated with the highest nontoxic concentrations of metal NPs, and the secretory activity of macrophages was determined. Macrophages cultivated in the presence of both untreated MSCs or MSCs preincubated with NPs produced significantly enhanced and comparable levels of various cytokines and growth factors. These results suggest that metal nanoparticles inhibit therapeutic properties of MSCs by a direct negative effect on their secretory activity, but MSCs cultivated in the presence of metal NPs have preserved the ability to stimulate cytokine and growth factor production by macrophages.

• Klíčová slova
• Cytokines, Cytotoxicity, Growth factors, Macrophages, Mesenchymal stem cells, Metal nanoparticles,
• MeSH
• cytokiny MeSH
• kovové nanočástice * MeSH
• mezenchymální kmenové buňky * MeSH
• myši MeSH
• vaskulární endoteliální růstový faktor A farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH
• titanium dioxide MeSH Prohlížeč
• vaskulární endoteliální růstový faktor A MeSH

Nanoparticles (NPs) have a wide use in various field of industry and in medicine, where they represent a promise for their antimicrobial effects. Simultaneous application of NPs and therapeutic stem cells can speed up tissue regeneration and improve healing process but there is a danger of negative impacts of NPs on stem cells. Therefore, we tested effects of four types of metal antimicrobial NPs on characteristics and function properties of mouse mesenchymal stem cells (MSCs) in vitro. All types of tested NPs, i.e. zinc oxide, silver, copper oxide and titanium dioxide, exerted negative effects on the expression of phenotypic markers, metabolic activity, differentiation potential, expression of genes for immunoregulatory molecules and on production of cytokines and growth factors by MSCs. However, there were apparent differences in the impact of individual types of NPs on tested characteristics and function properties of MSCs. The results showed that individual types of NPs influence the activity of MSCs, and thus the use of metal NPs during tissue regeneration and in combination with stem cell therapy should be well considered.

• Klíčová slova
• Cytokine production, Dose-dependent toxicity, Gene expression, Mesenchymal stem cells, Metal nanoparticles,
• MeSH
• antiinfekční látky * MeSH
• buněčná diferenciace MeSH
• hojení ran MeSH
• kovové nanočástice * toxicita   MeSH
• mezenchymální kmenové buňky * MeSH
• myši MeSH
• nanočástice * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antiinfekční látky * MeSH

An encounter of the developing immune system with an antigen results in the induction of immunological areactivity to this antigen. In the case of transplantation antigens, the application of allogeneic hematopoietic cells induces a state of neonatal transplantation tolerance. This tolerance depends on the establishment of cellular chimerism, when allogeneic cells survive in the neonatally treated recipient. Since mesenchymal stem/stromal cells (MSCs) have been shown to have low immunogenicity and often survive in allogeneic recipients, we attempted to use these cells for induction of transplantation tolerance. Newborn (less than 24 h old) C57BL/6 mice were injected intraperitoneally with 5 × 106 adipose tissue-derived MSCs isolated from allogeneic donors and the fate and survival of these cells were monitored. The impact of MSC application on the proportion of cell populations of the immune system and immunological reactivity was assessed. In addition, the survival of skin allografts in neonatally treated recipients was tested. We found that in vitro expanded MSCs did not survive in neonatal recipients, and the living MSCs were not detected few days after their application. Furthermore, there were no significant changes in the proportion of individual immune cell populations including CD4+ cell lineages, but we detected an apparent shift to the production of Th1 cytokines IL-2 and IFN-γ in neonatally treated mice. However, skin allografts in the MSC-treated recipients were promptly rejected. These results therefore show that in vitro expanded MSCs do not survive in neonatal recipients, but induce a cytokine imbalance without induction of transplantation tolerance.

• Klíčová slova
• Cytokines, Immunological reactivity, Mesenchymal stem cells, Newborn mice, Skin grafts, Transplantation tolerance,
• MeSH
• cytokiny MeSH
• interleukin-2 MeSH
• mezenchymální kmenové buňky * MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• novorozená zvířata MeSH
• transplantace mezenchymálních kmenových buněk * MeSH
• transplantační tolerance MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH
• interleukin-2 MeSH

Mesenchymal stem cells (MSCs) have the ability to migrate to the site of injury or inflammation, and to contribute to the healing process. Since patients treated with MSCs are often users of analgesic drugs, to relieve their uncomfortable pain associated with the tissue disorder, there is a possibility of negative effects of these drugs on the migration of endogenous and exogenous MSCs. Therefore, we tested the impact of acute and chronic treatment with morphine on the migration and organ distribution of exogenous adipose tissue-derived MSCs in mouse models. Firstly, we showed that the incubation of MSCs with morphine significantly reduced the expression of adhesive molecules CD44 (HCAM), CD54 (ICAM-1) and CD106 (VCAM-1) on MSCs. Using a model of systemic administration of MSCs labeled with vital dye PKH26 and by the application of flow cytometry to detect living CD45-PKH26+ cells, we found a decreased number of labeled MSCs in the lung, spleen and bone marrow, and a significantly increased number of MSCs in the liver of morphine-treated recipients. A skin allograft model was used to study the effects of morphine on the migration of exogenous MSCs to the superficial wound. Intraperitoneally administered MSCs migrated preferentially to the wound site, and this migration was significantly decreased in the morphine-treated recipients. The present results showed that morphine significantly influences the distribution of exogenous MSCs in the body, and decreases their migration to the site of injury.

• Klíčová slova
• Acute and chronic morphine treatment, Adhesive molecules, Cell migration, Mesenchymal stem cells, Skin graft model,
• MeSH
• antigeny CD44 MeSH
• cévní buněčněadhezivní molekula-1 MeSH
• hojení ran MeSH
• kůže zranění   MeSH
• mezenchymální kmenové buňky * cytologie   účinky léků   MeSH
• mezibuněčná adhezivní molekula-1 MeSH
• morfin * farmakologie   MeSH
• myši MeSH
• pohyb buněk * účinky léků   MeSH
• průtoková cytometrie MeSH
• rány a poranění MeSH
• tuková tkáň cytologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD44 MeSH
• cévní buněčněadhezivní molekula-1 MeSH
• Icam1 protein, mouse MeSH Prohlížeč
• mezibuněčná adhezivní molekula-1 MeSH
• morfin * MeSH

Mesenchymal stem cells (MSCs) represent a population of adult stem cells that have potent immunoregulatory, anti-inflammatory, and antiapoptotic properties. In addition, they have ability to migrate to the site of inflammation or injury, where they contribute to the regeneration and healing process. For these properties, MSCs have been used as therapeutic cells in several models, including treatment of damages or disorders of the ocular surface. If the damage of the ocular surface is extensive and involves a limbal region where limbal stem cell reside, MSC therapy has been proved as the effective treatment approach. Although the anti-inflammatory properties of MSCs have been well characterized, mechanisms of antiapoptotic action of MSCs are not well recognized. Using a chemically damaged cornea in a mouse model, we showed that the injury decreases expression of the gene for antiapoptotic molecule Bcl-2 and increases the expression of proapoptotic genes Bax and p53. These changes were attenuated by local transplantation of MSCs after corneal damage. The antiapoptotic effect of MSCs was tested in an in vitro model of co-cultivation of corneal explants with MSCs. The apoptosis of corneal cells in the explants was induced by proinflammatory cytokines and was significantly inhibited in the presence of MSCs. The antiapoptotic effect of MSCs was mediated by paracrine action, as confirmed by separation of the explants in inserts or by supernatants from MSCs. In addition, MSCs decreased the expression of genes for the molecules associated with endoplasmic reticulum stress Atf4, Bip, and p21, which are associated with apoptosis. The results show that MSCs inhibit the expression of proapoptotic genes and decrease the number of apoptotic cells in the damaged corneas, and this action might be one of the mechanisms of the therapeutic action of MSCs.

• Klíčová slova
• Bax, Bcl-2, antiapoptotic properties, cornea, mesenchymal stem cells, mouse model,
• MeSH
• apoptóza genetika   MeSH
• cyklooxygenasa 2 genetika   metabolismus   MeSH
• cytokiny genetika   metabolismus   MeSH
• hepatocytární růstový faktor genetika   metabolismus   MeSH
• keratitida genetika   metabolismus   patologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• modely nemocí na zvířatech * MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• poranění rohovky genetika   metabolismus   terapie   MeSH
• regulace genové exprese * MeSH
• rohovka metabolismus   MeSH
• transplantace mezenchymálních kmenových buněk metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cyklooxygenasa 2 MeSH
• cytokiny MeSH
• hepatocytární růstový faktor MeSH

The knowledge of mechanisms of regulation of IL-10 production by B cells remains still very limited. We show here that highly purified mouse B cells stimulated with LPS produce significant levels of IL-10, but Bregs in our model do not express detectable level of either Foxp3 or GATA-3. Nevertheless, IL-10 production by B cells is regulated by cytokines. In activated B cells, IL-10 production was significantly enhanced by IFN-γ and decreased in the presence of IL-4 or TGF-β. These findings are in sharp contrast with the observations in T cells, where IL-10 production correlates with GATA-3 or FoxP3 expression, and the cytokines regulate IL-10 production in a reverse manner than in activated B cells. These results thus show that the production of IL-10 by Bregs is regulated by cytokines independently of the expression of GATA-3 and FoxP3, which is clearly different from GATA-3-dependent IL-10 production by activated Th2 cells and FoxP3 expression in IL-10-producing Tregs.

• Klíčová slova
• B cell, Cytokine, FoxP3, GATA-3, IL-10 production, T cell,
• MeSH
• aktivace lymfocytů imunologie   MeSH
• faktor 1 indukovatelný hypoxií - podjednotka alfa imunologie   MeSH
• forkhead transkripční faktory metabolismus   MeSH
• interferon gama imunologie   MeSH
• interleukin-10 biosyntéza   MeSH
• interleukin-4 imunologie   MeSH
• kultivované buňky MeSH
• lipopolysacharidy imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• regulační B-lymfocyty imunologie   MeSH
• regulační T-lymfocyty imunologie   MeSH
• Th2 buňky imunologie   MeSH
• transformující růstový faktor beta imunologie   MeSH
• transkripční faktor GATA3 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• faktor 1 indukovatelný hypoxií - podjednotka alfa MeSH
• forkhead transkripční faktory MeSH
• Foxp3 protein, mouse MeSH Prohlížeč
• Gata3 protein, mouse MeSH Prohlížeč
• Hif1a protein, mouse MeSH Prohlížeč
• IFNG protein, mouse MeSH Prohlížeč
• IL10 protein, mouse MeSH Prohlížeč
• Il4 protein, mouse MeSH Prohlížeč
• interferon gama MeSH
• interleukin-10 MeSH
• interleukin-4 MeSH
• lipopolysacharidy MeSH
• transformující růstový faktor beta MeSH
• transkripční faktor GATA3 MeSH

Retinal degenerative disorders are characterized by a local upregulation of inflammatory factors, infiltration with cells of the immune system, a vascular dysfunction and by the damage of retinal cells. There is still a lack of treatment protocols for these diseases. Mesenchymal stem cell (MSC)-based therapy using immunoregulatory, regenerative and differentiating properties of MSCs offers a promising treatment option. In this study, we analyzed the immunomodulatory properties of mouse bone marrow-derived MSCs after their intravitreal delivery to the inflammatory environment in the eye, caused by the application of pro-inflammatory cytokines IL-1β, TNF-α and IFN-γ. The intravitreal administration of these cytokines induces an increased expression of pro-inflammatory molecules such as IL-1α, IL-6, inducible nitric oxide synthase, TNF-α and vascular endothelial growth factor in the retina. However, a significant decrease in the expression of genes for all these pro-inflammatory molecules was observed after the intravitreal injection of MSCs. We further showed that an increased infiltration of the retina with immune cells, mainly with macrophages, which was observed after pro-inflammatory cytokine application, was significantly reduced after the intravitreal application of MSCs. The similar immunosuppressive effects of MSCs were also demonstrated in vitro in cultures of cytokine-stimulated retinal explants and MSCs. Overall, the results show that intravitreal application of MSCs inhibits the early retinal inflammation caused by pro-inflammatory cytokines, and propose MSCs as a promising candidate for stem cell-based therapy of retinal degenerative diseases.

• Klíčová slova
• Cytokines, Immunomodulation, Inflammation, Mesenchymal stem cells, Retina,
• MeSH
• antivirové látky farmakologie   MeSH
• cytokiny metabolismus   MeSH
• imunomodulace účinky léků   imunologie   MeSH
• interferon gama farmakologie   MeSH
• interleukin-1beta farmakologie   MeSH
• mediátory zánětu farmakologie   MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• oxid dusnatý metabolismus   MeSH
• retina cytologie   účinky léků   imunologie   metabolismus   MeSH
• TNF-alfa farmakologie   MeSH
• zánět imunologie   metabolismus   patologie   prevence a kontrola   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antivirové látky MeSH
• cytokiny MeSH
• IL1B protein, mouse MeSH Prohlížeč
• interferon gama MeSH
• interleukin-1beta MeSH
• mediátory zánětu MeSH
• oxid dusnatý MeSH
• TNF-alfa MeSH

Retinal degenerative disorders, such as diabetic retinopathy, retinitis pigmentosa, age-related macular degeneration or glaucoma, represent the most common causes of loss of vision and blindness. In spite of intensive research, treatment options to prevent, stop or cure these diseases are limited. Newer therapeutic approaches are offered by stem cell-based therapy. To date, various types of stem cells have been evaluated in a range of models. Among them, mesenchymal stem/stromal cells (MSCs) derived from bone marrow or adipose tissue and used as autologous cells have been proposed to have the potential to attenuate the negative manifestations of retinal diseases. MSCs delivered to the vicinity of the diseased retina can exert local anti-inflammatory and repair-promoting/regenerative effects on retinal cells. However, MSCs also produce numerous factors that could have negative impacts on retinal regeneration. The secretory activity of MSCs is strongly influenced by the cytokine environment. Therefore, the interactions among the molecules produced by the diseased retina, cytokines secreted by inflammatory cells and factors produced by MSCs will decide the development and propagation of retinal diseases. Here we discuss the interactions among cytokines and other factors in the environment of the diseased retina treated by MSCs, and we present results supporting immunoregulatory and trophic roles of molecules secreted in the vicinity of the retina during MSC-based therapy.

• Klíčová slova
• Cytokines, Degenerative diseases, Growth factors, Mesenchymal stem cells, Retina, Stem cell therapy,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The inhalation or application of nanoparticles (NPs) has serious impacts on immunological reactivity. However, the effects of NPs on the immune system are influenced by numerous factors, which cause a high variability in the results. Here, mice were exposed to a three month continuous inhalation of copper oxide (CuO) NPs, and at different time intervals (3, 14, 42 and 93 days), the composition of cell populations of innate and adaptive immunity was evaluated in the spleen by flow cytometry. The ability of spleen cells from exposed and control mice to respond to stimulation with T- or B-cell mitogens by proliferation and by production of cytokines IL-2, IL-6, IL-10, IL-17 and IFN-γ was characterized. The results showed that the inhalation of CuO NPs predominantly affects the cells of innate immunity (changes in the proportion of eosinophils, neutrophils, macrophages and antigen-presenting cells) with a minimal effect on the percentage of T and B lymphocytes. However, the proliferative and secretory activity of T cells was already significantly enhanced after 3 days from the start of inhalation, decreased on day 14 and normalized at the later time intervals. There was no correlation between the impacts of NPs on the cells of innate and adaptive immunity. The results have shown that the inhalation of CuO NPs significantly alters the composition of cell populations of innate immunity and modulates the proliferation and production of cytokines by cells of the adaptive immune system. However, the immunomodulatory effects of inhaled NPs strongly depend on the time of inhalation.

• Klíčová slova
• CuO nanoparticles, adaptive immunity, immunoreactivity, inhalation, innate immunity,
• MeSH
• adaptivní imunita účinky léků   MeSH
• aplikace inhalační MeSH
• cytokiny biosyntéza   MeSH
• kinetika MeSH
• měď aplikace a dávkování   toxicita   MeSH
• modely u zvířat MeSH
• myši inbrední ICR MeSH
• myši MeSH
• nanočástice toxicita   MeSH
• přirozená imunita účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cupric oxide MeSH Prohlížeč
• cytokiny MeSH
• měď MeSH