PURPOSE: The aim of this work was to demonstrate an immunostimulatory and adjuvant effect of new apyrogenic lipophilic derivatives of norAbuMDP and norAbuGMDP formulated in nanoliposomes. METHODS: Nanoliposomes and metallochelating nanoliposomes were prepared by lipid film hydration and extrusion methods. The structure of the liposomal formulation was studied by electron microscopy, AF microscopy, and dynamic light scattering. Sublethal and lethal γ-irradiation mice models were used to demonstrate stimulation of innate immune system. Recombinant Hsp90 antigen (Candida albicans) bound onto metallochelating nanoliposomes was used for immunisation of mice to demonstrate adjuvant activities of tested compounds. RESULTS: Safety and stimulation of innate and adaptive immunity were demonstrated on rabbits and mice. The liposomal formulation of norAbuMDP/GMDP was apyrogenic in rabbit test and lacking any side effect in vivo. Recovery of bone marrow after sublethal γ-irradiation as well as increased survival of mice after lethal irradiation was demonstrated. Enhancement of specific immune response was demonstrated for some derivatives incorporated in metallochelating nanoliposomes with recombinant Hsp90 protein antigen. CONCLUSIONS: Liposomal formulations of new lipophilic derivatives of norAbuMDP/GMDP proved themselves as promising adjuvants for recombinant vaccines as well as immunomodulators for stimulation of innate immunity and bone-marrow recovery after chemo/radio therapy of cancer.

• MeSH
• acetylmuramyl-alanyl-isoglutamin aplikace a dávkování   analogy a deriváty   chemie   farmakologie   terapeutické užití   MeSH
• adaptivní imunita účinky léků   MeSH
• adjuvancia imunologická aplikace a dávkování   chemie   farmakologie   terapeutické užití   MeSH
• analýza přežití MeSH
• antigeny fungální imunologie   MeSH
• experimentální radiační poranění imunologie   prevence a kontrola   MeSH
• králíci MeSH
• liposomy MeSH
• mikroskopie atomárních sil MeSH
• mikroskopie elektronová rastrovací MeSH
• molekulární struktura MeSH
• myši inbrední ICR MeSH
• myši MeSH
• nanočástice MeSH
• nosiče léků chemie   MeSH
• přirozená imunita účinky léků   MeSH
• proteiny tepelného šoku HSP90 imunologie   MeSH
• protilátky fungální krev   MeSH
• rekombinantní proteiny imunologie   MeSH
• transmisní elektronová mikroskopie MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

PURPOSE: Research areas of 'post-exposure treatment' and 'cytokines and growth factors' have top priority among studies aimed at radiological nuclear threat countermeasures. The experiments were aimed at testing the ability of N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA), an adenosine A(3) receptor agonist, to modulate hematopoiesis in sublethally irradiated mice, when administered alone or in a combination with granulocyte colony-stimulating factor (G-CSF) in a two-day post-irradiation treatment regimen. MATERIALS AND METHODS: A complete analysis of hematopoiesis including determination of numbers of bone marrow hematopoietic progenitor and precursor cells, as well as of numbers of peripheral blood cells, was performed. The outcomes of the treatment were assessed at days 3 to 22 after irradiation. RESULTS: IB-MECA alone has been found to induce a significant elevation of numbers of bone marrow granulocyte-macrophage progenitor cells (GM-CFC) and peripheral blood neutrophils. IB-MECA given concomitantly with G-CSF increased significantly bone marrow GM-CFC and erythroid progenitor cells (BFU-E) in comparison with the controls and with animals administered each of the drugs alone. CONCLUSIONS: The findings suggest the ability of IB-MECA to stimulate hematopoiesis and to support the hematopoiesis-stimulating effects of G-CSF in sublethally irradiated mice.

• MeSH
• adenosin analogy a deriváty   farmakologie   MeSH
• celotělové ozáření MeSH
• erytroidní prekurzorové buňky účinky záření   MeSH
• faktor stimulující kolonie granulocytů krev   farmakologie   MeSH
• hematopoéza účinky léků   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• receptor adenosinový A3 fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Expression of mRNA for adenosine receptor subtypes A(1), A(2a), A(2b), and A(3) in normal and lipopolysaccharide (LPS)-activated murine RAW 264.7 macrophages has been investigated using the method of quantitative real-time polymerase chain reaction. The results have shown a very low, unquantifiable expression of adenosine A(1) receptor mRNA in both normal and LPS-activated macrophages. The other three adenosine receptor mRNAs have been found to be expressed at various but always quantifiable levels. Activation of the macrophages by LPS induced upregulation of the expression of adenosine receptor A(2a) and A(2b) mRNA, whereas the expression of adenosine receptor A(3) mRNA was downregulated. Unstimulated macrophages exhibited a high expression of the A(2b) adenosine receptor mRNA. The findings are discussed from the point of view of the antiinflammatory and hematopoiesis-stimulating roles of the adenosine receptor signaling.

• MeSH
• aktivace makrofágů genetika   účinky léků   MeSH
• buněčné linie MeSH
• časové faktory MeSH
• financování organizované MeSH
• hematopoéza genetika   účinky léků   MeSH
• lipopolysacharidy farmakologie   MeSH
• makrofágy metabolismus   účinky léků   MeSH
• messenger RNA metabolismus   MeSH
• myši MeSH
• polymerázová řetězová reakce MeSH
• purinergní receptory P1 genetika   účinky léků   MeSH
• receptor adenosinový A1 genetika   MeSH
• receptor adenosinový A2A genetika   MeSH
• receptor adenosinový A2B genetika   MeSH
• receptor adenosinový A3 genetika   MeSH
• regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH

Adenosine A(3) receptor agonist N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA) has been tested from the point of view of potentiating the effects of hematopoietic growth factors interleukin-3 (IL-3), stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF) on the growth of hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) in suspension of normal mouse bone marrow cells in vitro. IB-MECA alone induced no GM-CFC growth. Significant elevation of numbers of GM-CFC evoked by the combinations of IB-MECA with IL-3, SCF, or GM-CSF as compared with these growth factors alone has been noted. Combination of IB-MECA with G-CSF did not induce significantly higher numbers of GM-CFC in comparison with G-CSF alone. Joint action of three drugs, namely of IB-MECA + IL-3 + GM-CSF, produced significantly higher numbers of GM-CFC in comparison with the combinations of IB-MECA + IL-3, IB-MECA + GM-CSF, or IL-3 + GM-CSF. These results give evidence of a significant role of selective activation of adenosine A(3) receptors in stimulation of the growth of granulocyte/ macrophage hematopoietic progenitor cells.

• MeSH
• adenosin analogy a deriváty   farmakologie   MeSH
• buněčné dělení účinky léků   MeSH
• faktor stimulující granulocyto-makrofágové kolonie farmakologie   MeSH
• financování organizované MeSH
• granulocyty cytologie   MeSH
• hematopoetické kmenové buňky cytologie   metabolismus   účinky léků   MeSH
• hepatocytární růstový faktor farmakologie   MeSH
• inbrední kmeny myší MeSH
• interleukin-3 farmakologie   MeSH
• kultivované buňky MeSH
• makrofágy cytologie   MeSH
• myši MeSH
• receptor adenosinový A3 metabolismus   MeSH
• synergismus léků MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH

The vitamin E analogue alpha-tocopheryl succinate (alpha-TOS) is an efficient anti-cancer drug. Improved efficacy was achieved through the synthesis of alpha-tocopheryl maleamide (alpha-TAM), an esterase-resistant analogue of alpha-tocopheryl maleate. In vitro tests demonstrated significantly higher cytotoxicity of alpha-TAM towards cancer cells (MCF-7, B16F10) compared to alpha-TOS and other analogues prone to esterase-catalyzed hydrolysis. However, in vitro models demonstrated that alpha-TAM was cytotoxic to non-malignant cells (e.g. lymphocytes and bone marrow progenitors). Thus we developed lyophilized liposomal formulations of both alpha-TOS and alpha-TAM to solve the problem with cytotoxicity of free alpha-TAM (neurotoxicity and anaphylaxis), as well as the low solubility of both drugs. Remarkably, neither acute toxicity nor immunotoxicity implicated by in vitro tests was detected in vivo after application of liposomal alpha-TAM, which significantly reduced the growth of cancer cells in hollow fiber implants. Moreover, liposomal formulation of alpha-TAM and alpha-TOS each prevented the growth of tumours in transgenic FVB/N c-neu mice bearing spontaneous breast carcinomas. Liposomal formulation of alpha-TAM demonstrated anti-cancer activity at levels 10-fold lower than those of alpha-TOS. Thus, the liposomal formulation of alpha-TAM preserved its strong anti-cancer efficacy while eliminating the in vivo toxicity found of the free drug applied in DMSO. Liposome-based targeted delivery systems for analogues of vitamin E are of interest for further development of efficient and safe drug formulations for clinical trials.

• MeSH
• alfa-tokoferol analogy a deriváty   aplikace a dávkování   farmakologie   MeSH
• antitumorózní látky aplikace a dávkování   farmakologie   MeSH
• chemie farmaceutická MeSH
• lidé MeSH
• liposomy MeSH
• maleimidy aplikace a dávkování   farmakologie   MeSH
• melanom experimentální farmakoterapie   patologie   MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši transgenní MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory prsu farmakoterapie   patologie   MeSH
• polyethylenglykoly aplikace a dávkování   farmakologie   MeSH
• vitamin E analogy a deriváty   aplikace a dávkování   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Meloxicam, a selective inhibitor of cyclooxygenase 2, was tested to determine its ability to modulate hematopoiesis and to influence survival of mid-lethally gamma-irradiated mice. A single dose of meloxicam (20 mg/kg) administered to mice intraperitoneally 1 h before irradiation was shown to enhance serum levels of granulocyte colony-stimulating factor (G-CSF) during the first 24 h after irradiation, to elevate numbers of granulocytic precursor cells in bone marrow and granulocyte counts in peripheral blood on day 10 after irradiation, and to increase 30-day survival of these mice. The results provide new evidence for the protective ability of meloxicam administration to mice irradiated with mid-lethal doses and contribute to the understanding of the mechanisms of this meloxicam action by drawing attention to the possible role of increased endogenous G-CSF production.

• MeSH
• faktor stimulující kolonie granulocytů biosyntéza   MeSH
• financování organizované MeSH
• hematopoéza účinky záření   MeSH
• inhibitory cyklooxygenasy 2 farmakologie   MeSH
• myši MeSH
• radioprotektivní látky farmakologie   MeSH
• thiaziny farmakologie   MeSH
• thiazoly farmakologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH

Hematopoiesis-modulating action of meloxicam, a cyclooxyge-nase-2 inhibitor, has been evaluated in mice. Increased serum level of granulocyte colony-stimulating factor (G-CSF) after meloxicam administration has been found in sublethally gamma-irradiated animals. In further experiments hematopoiesis-stimulating effects of meloxicam and G-CSF given alone or in combination have been investigated. Granulocyte/macrophage progenitor cells counts were used to monitor these effects. Meloxicam and exogenous G-CSF did not act synergistically when given in combination, but could be mutually substituted during their repeated administration. The results suggest a promising possibility of using meloxicam as an auxiliary drug reducing the high costs of G-CSF therapy of myelosuppression.

• MeSH
• buněčná diferenciace MeSH
• celotělové ozáření MeSH
• faktor stimulující kolonie granulocytů aplikace a dávkování   krev   účinky léků   MeSH
• granulocyty cytologie   účinky léků   účinky záření   MeSH
• hematopoéza účinky léků   MeSH
• inhibitory cyklooxygenasy 2 farmakologie   MeSH
• kombinovaná farmakoterapie MeSH
• kostní dřeň účinky léků   účinky záření   MeSH
• lékové interakce MeSH
• leukopenie prevence a kontrola   MeSH
• myši inbrední CBA MeSH
• myši MeSH
• thiaziny farmakologie   MeSH
• thiazoly farmakologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH

Two adenosine receptor agonists, N6-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA) and N6-cyclopentyladenosine (CPA), which selectively activate adenosine A3 and A1 receptors, respectively, were tested for their ability to influence proliferation of granulocytic and erythroid cells in femoral bone marrow of mice using morphological criteria. Agonists were given intraperitoneally to mice in repeated isomolar doses of 200 nmol/kg. Three variants of experiments were performed to investigate the action of the agonists under normal resting state of mice and in phases of cell depletion and subsequent regeneration after treatment with the cytotoxic drug 5-fluorouracil. In the case of granulopoiesis, IB-MECA 1) increased by a moderate but significant level proliferation of cells under normal resting state; 2) strongly increased proliferation of cells in the cell depletion phase; but 3) did not influence cell proliferation in the regeneration phase. CPA did not influence cell proliferation under normal resting state and in the cell depletion phase, but strongly suppressed the overshooting cell proliferation in the regeneration phase. The stimulatory effect of IB-MECA on cell proliferation of erythroid cells was observed only when this agonist was administered during the cell depletion phase. CPA did not modulate erythroid proliferation in any of the functional states investigated, probably due to the lower demand for cell production as compared with granulopoiesis. The results indicate opposite effects of the two adenosine receptor agonists on proliferation of hematopoietic cells and suggest the plasticity and homeostatic role of the adenosine receptor expression.

• MeSH
• adenosin analogy a deriváty   farmakologie   MeSH
• agonisté adenosinového receptoru A1 MeSH
• agonisté adenosinového receptoru A3 MeSH
• buňky kostní dřeně cytologie   metabolismus   účinky léků   MeSH
• erytroidní buňky cytologie   metabolismus   účinky léků   MeSH
• financování organizované MeSH
• granulocyty cytologie   metabolismus   účinky léků   MeSH
• hematopoetické kmenové buňky cytologie   metabolismus   účinky léků   MeSH
• homeostáza fyziologie   MeSH
• myši inbrední C57BL MeSH
• myši inbrední CBA MeSH
• myši MeSH
• neurotransmiterové látky MeSH
• proliferace buněk účinky léků   MeSH
• receptor adenosinový A1 MeSH
• receptor adenosinový A3 metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH

IMUNOR, a low-molecular weight (< 12 kD) ultrafiltered pig leukocyte extract, has been previously found to have significant stimulatory effects on murine hematopoiesis supressed by ionizing radiation or cytotoxic drugs. This communication shows data on the mechanisms of these effects. Using ELISA assay, significantly increased levels of granulocyte colony-stimulating factor (G-CSF) and interleukin-6 (IL-6) were observed. On the contrary, no detectable levels of granulocyte-macrophage colony-stimulating factor (GM-CFC) and interleukin-3 (IL-3) have been found in blood serum of IMUNOR-treated mice. Incubation of the serum from IMUNOR-treated mice with antibodies against G-CSF caused abrogation of the ability of the sera to stimulate in vitro growth of colonies originating from granulocyte-macrophage progenitor cells (GM-CFC). In contrast, incubation of the serum with antibodies against IL-6 did not change its colony-stimulating activity. It may be inferred from these findings that G-CSF is probably the main cytokine responsible for the granulopoiesis-stimulating effects of IMUNOR. When the serum from IMUNOR-treated mice with G-CSF inactivated by anti-G-CSF antibodies (but with elevated IL-6) was added to cultures of bone marrow cells together with a suboptimum concentration of IL-3, a significant increase in the numbers of GM-CFC colonies was found. Moreover, conjoint inactivation of G-CSF and IL-6 significantly decreased the numbers of GM-CFC colonies in comparison with those observed when only G-CSF was inactivated. This observation strongly suggests that though IMUNOR-induced IL-6 is not able to induce the growth of GM-CFC colonies alone, it is able to potentiate the hematopoiesis-stimulating effect of IL-3. These findings represent a new knowledge concerning the hematopoiesis-stimulating action of IMUNOR, a promising immunomodulatory agent.

• MeSH
• antisérum farmakologie   MeSH
• chemická stimulace MeSH
• ELISA MeSH
• faktor stimulující kolonie granulocytů antagonisté a inhibitory   fyziologie   MeSH
• financování organizované MeSH
• granulocyty fyziologie   MeSH
• hematopoéza účinky léků   MeSH
• interleukin-3 antagonisté a inhibitory   MeSH
• interleukin-6 antagonisté a inhibitory   fyziologie   MeSH
• kostní dřeň metabolismus   účinky léků   MeSH
• leukocyty fyziologie   MeSH
• monoklonální protilátky farmakologie   MeSH
• myši MeSH
• prasata MeSH
• protilátky blokující farmakologie   MeSH
• tkáňové extrakty farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH

A low-molecular-weight (<12 kDa) ultrafiltered pig leukocyte extract, IMUNOR, was tested in experiments in vitro on non-stimulated and lipopolysaccharide (LPS)-stimulated murine RAW 264.7 macrophages in order to assess modulation of nitric oxide (NO) production (measured indirectly as the concentration of nitrites), hematopoiesis-stimulating activity of the supernatant of the macrophage cells (ascertained by counting cell colonies growing from progenitor cells for granulocytes and macrophages (GM-CFC) in vitro), and the release of hematopoiesis-stimulating cytokines. No hematopoiesis-stimulating activity and cytokine or NO production were found in the supernatant of non-stimulated macrophages. It was found that IMUNOR does not influence this status. Supernatant of LPS-stimulated macrophages was characterized by hematopoiesis-stimulating activity, as well as by the presence of nitrites, interleukin-6 (IL-6), and granulocyte colony-stimulating factor (G-CSF). A key role in the hematopoiesis-stimulating activity of the supernatant of LPS-stimulated macrophages could be ascribed to G-CSF since the formation of the colonies could be abrogated nearly completely by monoclonal antibodies against G-CSF. IMUNOR was found to suppress all the mentioned manifestations of the LPS-activated macrophages. When considering these results together with those from our previous in vivo study revealing stimulatory effects of IMUNOR on radiation-suppressed hematopoiesis, a hypothesis may be formulated which postulates a homeostatic role of IMUNOR, consisting in stimulation of impaired immune and hematopoietic systems but also in cutting back the production of proinflammatory mediators in cases of overstimulation which threats with undesirable consequences.

• MeSH
• buněčné linie MeSH
• buňky kostní dřeně cytologie   účinky léků   MeSH
• cytokiny metabolismus   MeSH
• faktor stimulující kolonie granulocytů farmakologie   metabolismus   MeSH
• financování organizované MeSH
• hematopoéza účinky léků   MeSH
• imunologické faktory farmakologie   MeSH
• inbrední kmeny myší MeSH
• kultivované buňky MeSH
• lipopolysacharidy MeSH
• makrofágy metabolismus   účinky léků   MeSH
• myši MeSH
• oxid dusnatý metabolismus   MeSH
• prasata MeSH
• tkáňové extrakty MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH