M Colić

University of Belgrade, Beograd, Central Serbia, Serbia

Are you M Colić?

Claim your profile

Publications (220)286.05 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Introduction: Thymic dendritic cells (TDCs) are crucial cells in process of negative selection of auto reactive thymocytes. Antigenic peptides presented by TDCs could be derived from their endogenous antigens, or exogenous antigens uptaken from surrounding cells by endocytosis and phagocytosis. Considering our previous findings, that GM-CSF in vitro significantly modulate accessory function of TDCs, in this study we tested the influence of GM-CSF on antigen internalisation by CD11b- and CD11b+TDCs subsets. Material and methods: TDCs were isolated from the thymus of male AO rats, using the density centrifugation and TDCs subsets were immunomagnetic separated according to the expression of CD11b molecules. The endocytic and phagocytic activity were analyzed as uptake of dextran-FITC and CMFD labeled apoptotic thymocytes, respectively. Endocytic and phagocytic activity were evaluated by flow cytometry after staining TDCs with anti-MHC-class II (OX6-PE mAb) Antigen internalisation was quantified by the presence of double positive cells (OX-6-PE+/FITC-dextran+ and OX-6-PE+/CMFD+). Results: CD11b-TDCs were more potent in FITC-dextran uptake than CD11b+TDCs, but there was no difference in their ability to phagocyte the apoptotic thymocytes. When these TDCs were cultured with GM-CSF, a significant increase in endocytic and phagocytic activity was observed only in CD11b+TDCs subset. We further analyzed antigen uptake at different maturation stages of both TDCs subsets. In contrast to endocytosis, mature (MHC class-IIhigh) TDCs showed higher phagocytic activity than immature (MHC class-IIlow) TDCs. Stimulatory effect of GM-CSF on phagocytic activity was manifested on CD11b+ but not CD11b-TDCs independently on maturation stage. Conclusion: CD11b-TDCs could be more efficient in uptake of soluble antigens whereas both TDCs subsets are equally efficient in phagocytosis of apoptotic cells. The significant increasing in endocytic capacity of CD11b+TDCs under the influence of GM-CSF suggests the role of this cytokine as potentially selective regulator of antigen internalization in the thymus.
    No preview · Article · Dec 2015
  • Source
    Milan Markovic · Sergej Tomic · Jelena Djokic · Miodrag Colic
    [Show abstract] [Hide abstract]
    ABSTRACT: The pathophysiology of periapical lesions (PLs) is under control of pro-inflammatory and anti-inflammatory (mainly immunoregulatory) cytokines. We have recently established mesenchymal stem cells (MSCs) from PLs and showed their suppressive effects on the production of proinflammatory cytokines from PLs inflammatory cells (ICs). In this work we studied the production of interleukin (IL)-10, IL-27 and transforming growth factor (TGF)-β, by PL-ICs in direct or indirect contacts with PL-MSCs. PL-ICs, which were isolated from four different asymptomatic PLs, predominantly composed of lymphocytes, followed by neutrophil granulocytes, macrophages and plasma cells. PLMSCs, expressing typical MSC markers, were co-cultivated with PL-ICs at 1:10 ratio, either in direct contact or in a transwell-system, for 24 hours. The levels of cytokines in cell-culture supernatants were tested by ELISA. The results showed that PL-MSCs up-regulated the production of all three immunoregulatory cytokines by PL-ICs. PL-MSCs stimulated the production of IL-10 and IL-27 via soluble factors, whereas the up-regulation of TGF-β required direct cell-to-cell contacts. In conclusion, our results showed for the first time the involvement of PL-MSCs in restriction of inflammation in PLs by up-regulation of immunoregulatory cytokines.
    Full-text · Article · Oct 2015 · Acta Facultatis Medicae Naissensis
  • [Show abstract] [Hide abstract]
    ABSTRACT: Considering that gut mucosal and tumor tissue represents immunologycally tolerogenic microenvironments, we wonder whether pretreatment of monocyte- derived dendritic cells (MoDCs) with colorectal carcinoma conditioned media (CCM-MoDCs) or CM from normal colon mucosa tissue (NCM-MoDCs) may influence their functional response to TLR-3 and TLR-7 ligation. Tumour tissue as well as normal colon mucosa tissue was collected from colorectal carcinoma patients, whereas MoDCs were generated from healthy donors. MoDCs were cultivated with CCM, NCM or without CM (control MoDCs) for 4h and additionally for 18h with polyI:C and 7-thia-8-oxoguanosine (TLR-3 and TLR-7 agonists, respectively). CCM-MoDCs and NCM-MoDCs up-regulated CD83, CD86, HLA-DR and increased secretion of TNF-α, IL-6, IL-10 and IL-23 compared to control MoDCs. Both, CCM-MoDCs and NCM-MoDCs showed similar phenotype features. CCM-MoDCs secreted lower levels of TNF-α and IL-23 but higher levels of IL-10 compared to NCM-MoDCs. Additional stimulation with TLR agonsts induced higher production of IL-12p70 and very strong production of IL-10 and IL-23 by both CM treated MoDCs and blocked production of TNF-α by CCM-MoDCs. Phenotipicaly, both MoDCs types slightly up-regulated HLA-DR and CD86, and almost completely down-regulated CD83. CCM-MoDCs and NCM-MoDCs responded to TLR simulation with very high production of IL-10 and IL-23 and significantly lower allostimulatory and Th1-polarization capacity compared to control MoDCs. Our findings suggests that CCM- and NCM-educated MoDCs can not be in vitro reverted into potent immunostimulatory DCs using TLR-3 and TLR-7 agonists, implying that in vivo application of these agonists could impair Th1 polarization capacity of tumor infiltrated DCs.
    No preview · Conference Paper · Sep 2015
  • [Show abstract] [Hide abstract]
    ABSTRACT: Introduction: Langerhans cells (LCs) represent a subpopulation of dendritic cells which initiate immune responses against the antigens from epidermis and mucoses. We have studied effects of Toll-like receptor 3 (TLR-3) and Dectin-1 agonists, (polyinosinic:polycytidylic acid (PolyI:C) and curdlan, respectively) on maturation of their in vitro counterparts, monocyte derived LCs (MoLCs), after ligation of CD40 molecule. Methods: Immature MoLCs were generated in the presence of GM-CSF, IL-4 and TGF-β1 and then treated with PolyI:C, curdlan or their combination for 24h. After that, cells were washed and additionally stimulated with soluble CD40L for another 24h. Results: Both single and combined treatments of MoLCs followed by ligation of CD40 molecule elicited phenotypic maturation of MoLCs and had different effect on their Th polarization capability. Poly I:C-treated MoLCs after ligation of CD40 molecule down-regulated both the production of IFN-γ and IL-17, while curdlan-treated MoLCs down-regulated production of IL-17 by alloreactive CD4+ T-cells compared to non CD40L-treated cells. Ligation of CD40 molecule on Poly I:C+curdlan-treated MoLCs was manifested by higher production of TNF-α, IL-1β, IL-23 and IL-27. These cells showed higher ability to induce production of IFN-γ, IL-2 and IL-17 by alloreactive CD4+ T-cells then curdlan- or Poly(I:C)-treated MoLCs. The IFN-γ/IL-4 ratio was higher in the cocultures with Poly I:C+curdlan-treated MoLCs suggesting their stronger Th1 polarization capability than separately-treated MoLC. Conclusion: Combined stimulation of MoLCs with TLR-3 and Dectin-1 agonists potentiate their Th1 polarization capability suggesting the role of these molecules for preparation of Th1 polarized immunostimulatory LCs in vitro.
    No preview · Conference Paper · Sep 2015
  • [Show abstract] [Hide abstract]
    ABSTRACT: Introduction: Functional impairment of dendritic cells (DCs) is one of important mechanisms in tumor-mediated immune response subversion. Considering that tumor tissue conditioned media (TCM) could represent suitable source of tumor antigens we wondered whether tumor extracts can be used for in vitro preparation of DCs with stable functional properties. Materials and Methods: Different concentration of TCM (50%, 25%, 10%, 5%) from colorectal carcinoma patients were added to mitogen-stimulated peripheral blood cells (mPBMNCs) and monocyte-derived DCs (MoDCs) of healthy donors. After 18h MoDCs were washed analyzed for their viability and accessory function. Results: Assessment of proliferative capacity of mPBMNCs revealed that 50%TCM almost completely inhibited their proliferation. However, recovering of mPBMNC proliferative capacity was inversely correlated with TCM concentration. Similar was observed in culture of MoDCs with allogeneic T-cells. MoDCs treated with the lowest TCM concentration (5%) showed significantly higher allostimulatory capacity then control cells. Analysis of viability revealed that 50%TCM induces more than 75% necrosis of MoDCs, whereas 25-5%CCM induces necrosis at the level of control cells. Analysis of CD40, CD83 and CD86 expression showed that all molecules were up-regulated by TCM at concentration lower than 25%. Their highest up-regulation was in the 5%CCM-treated MoDCs and expression of these molecules was similar as in TLR-3/7-treated MoDCs. Conclusions: The finding that TCM at low concentration can induce DCs-maturation in a similar extent as TLR-agonists, suggesting possible application of TCM in maturation protocols where TCM at the same time could be the source of tumor antigens and DCs-maturation factors.
    No preview · Conference Paper · Sep 2015
  • Source
    Dragan Mikic · Sasa Vasilijic · Milica Cucuz · Miodrag Colic
    [Show abstract] [Hide abstract]
    ABSTRACT: Background/Aim. The goal of modern clinical and experimental researches in the field of sepsis is to find one or more sensitive parameters that could predict the severity of sepsis and its outcome. In this study we investigated and compared the relationship of initial soluble Fas (sFas) plasma levels as well as Acute Physiology, Age and Chronic Health Evaluation II (APACHE II) score in 58 septic patients with severity and outcome of sepsis. Methods. The diagnosis and assessment of disease severity was performed on the same day, based on clinical and laboratory parameters. The blood samples were used for monitoring of laboratory standard parameters necessary for the diagnosis of sepsis, organ dysfunction and assessment of disease severity, as well as for determination of levels of sFas. According to consensus criteria, patients were divided into those with sepsis (n = 16), severe sepsis (n = 30) or septic shock (n = 12), those. with (n = 26) and without (n = 32) multiple organ dysfunction syndrome (MODS), and survivors (n = 45) and non-survivors (n = 13). Results. Plasma sFas level (9.7 +/- 10.1; 0-44.2 U/mL) was elevated in 54.4% of patients. All the patients with septic shock, 76.9% of the patients with MODS and 84.6% patients who died had elevated sFas level. We observed a strong positive correlation between sFas and APACHE II score (p < 0.001). The level of sFas was significantly higher in patients with septic shock compared to normotensive patients (p < 0.001), patients with MODS compared to those without MODS (p < 0.001) and survivors compared to nonsurvivors (p < 0.01). Conclusions. Our results suggest that initial sFas plasma levels in patients with sepsis correlated with the values of APACHE II score and separated very well the patients with septic shock versus the normotensive patients, the patients with and without MODS, and survivors versus non-survivors.
    Preview · Article · Jul 2015 · Vojnosanitetski pregled. Military-medical and pharmaceutical review
  • [Show abstract] [Hide abstract]
    ABSTRACT: Hashimoto thyroiditis (HT) is the most frequent thyroid autoimmune disease, while papillary thyroid cancer (PTC) is one of the most common endocrine malignancies. A few patients with HT also develop PTC. The aim of this study was to analyze cytokine profiles in patients with PTC accompanied with autoimmune HT in comparison with those in patients with PTC alone or HT alone and healthy subjects. Cytokine levels were determined in supernatants obtained from phytohemagglutinin (PHA)-stimulated whole blood cultures in vitro. The concentrations of selected cytokines: Th1-interferon gamma (IFN-γ); Th2-interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 10 (IL-10) and interleukin 13 (IL-13); Th9-interleukin 9 (IL-9); and Th17-interleukin 17 (IL-17A) were measured using multiplex cytokine detection systems for human Th1/Th2/Th9/Th17/Th22. We found that PTC patients with HT produced significantly higher concentrations of IL-4, IL-6, IL-9, IL-13 and IFN-γ than PTC patients without HT. In conclusion, autoimmune HT affects the cytokine profile of patients with PTC by stimulating secretion of Th1/Th2/Th9 types of cytokines. Th1/Th2 cytokine ratios in PTC patients with associated autoimmune HT indicate a marked shift toward Th2 immunity.
    No preview · Article · May 2015 · Cancer Immunology and Immunotherapy
  • [Show abstract] [Hide abstract]
    ABSTRACT: Mesenchymal stem cells (MSCs) increasingly attract considerable attention for cell-based therapies of damaged tissues and chronic inflammatory diseases. However, the mechanisms by which MSCs interact with innate phagocytic immune cells, such as dendritic cells (DCs), macrophages and neutrophils during inflammation, remain poorly investigated. Therefore, we investigated the differentiation potential of MSCs isolated from the dental periapical lesion (PL), chronic inflammatory granulomatous tissue, and their effect on phagocytic cells in vitro. In addition to the self-renewal capacity and expression of markers characteristic for MSCs, PL-MSCs demonstrated multi-differential potential with the preference to osteoblast differentiation. These cells produce spontaneously high amounts of IL-6 and IL-8 and moderate levels of TGF-β and IL-27. We showed that PL-MSCs inhibited both spontaneous and fMLP-induced apoptosis of neutrophils and potentiated survival of these cells triggered by LPS. Additionally, PL-MSCs suppressed differentiation of DCs from monocytes in the presence of GM-CSF and IL-4, as judged by significantly higher expression of CD14 and lower expression of CD1a on DCs. PL-MSCs induce differentiation of monocytes to regulatory macrophages, characterized by lower expression of CD169, higher expression of CD14, and the ability of these cells to produce high levels of IL-6 upon stimulation with LPS and IFN-γ. All these results correlated with the ability of PL-MSCs to down-regulate the production of IL-1β, IL-6, IL-8 and TNF-α by inflammatory cells from PL. These in vitro findings correlated with frequent contacts of DCs (HLA-DR+CD209+ cells), macrophages (CD68+ cells) and neutrophils (MPO+CD15+ cells) with perivascular PL-MSCs (CD146+STRO1+ cells) in situ as revealed by confocal microscopy. These results suggest that PL-MSCs promote immunoregulatory mechanisms by altering the functions of key innate phagocytic immune cells.
    No preview · Conference Paper · Oct 2014
  • [Show abstract] [Hide abstract]
    ABSTRACT: Background & objectives: Aluminum (Al) toxicity is closely linked to the pathogenesis of Alzheimer's disease (AD). This experimental study was aimed to investigate the active avoidance behaviour of rats after intrahippocampal injection of Al, and biochemical and immunohistochemical changes in three bilateral brain structures namely, forebrain cortex (FBCx), hippocampus and basal forebrain (BF). Methods: Seven days after intra-hippocampal (CA1 sector) injection of AlCl3 into adult male Wistar rats they were subjected to two-way active avoidance (AA) tests over five consecutive days. Control rats were treated with 0.9% w/v saline. The animals were decapitated on the day 12 post-injection. The activities of acetylcholinesterase (AChE) and glucose-6-phosphate dehydrogenase (G6PDH) were measured in the FBCx, hippocampus and BF. Immunohistochemical staining was performed for transferrin receptors, amyloid β and tau protein. Results: The activities of both AChE and G6PDH were found to be decreased bilaterally in the FBCx, hippocampus and basal forebrain compared to those of control rats. The number of correct AA responses was reduced by AlCl3 treatment. G6PDH administered prior to AlCl3 resulted in a reversal of the effects of AlCl3 on both biochemical and behavioural parameters. Strong immunohistochemical staining of transferrin receptors was found bilaterally in the FBCx and the hippocampus in all three study groups. In addition, very strong amyloid β staining was detected bilaterally in all structures in AlCl3-treated rats but was moderate in G6PDH/AlCl3-treated rats. Strong tau staining was noted bilaterally in AlCl3-treated rats. In contrast, tau staining was only moderate in G6PDH/AlCl3-treated rats. Interpretation & conclusions: Our findings indicated that the G6PDH alleviated the signs of behavioural and biochemical effects of AlCl3-treatment suggesting its involvement in the pathogenesis of Al neurotoxicity and its potential therapeutic benefit. The present model could serve as a useful tool in AD investigations.
    No preview · Article · Jun 2014 · The Indian Journal of Medical Research
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Gold nanoparticles (GNPs) are claimed as outstanding biomedical tools for cancer diagnostics and photo-thermal therapy, but without enough evidence on their potentially adverse immunological effects. Using a model of human dendritic cells (DCs), we showed that 10 nm- and 50 nm-sized GNPs (GNP10 and GNP50, respectively) were internalized predominantly via dynamin-dependent mechanisms, and they both impaired LPS-induced maturation and allostimulatory capacity of DCs, although the effect of GNP10 was more prominent. However, GNP10 inhibited LPS-induced production of IL-12p70 by DCs, and potentiated their Th2 polarization capacity, while GNP50 promoted Th17 polarization. Such effects of GNP10 correlated with a stronger inhibition of LPS-induced changes in Ca2+ oscillations, their higher number per DC, and more frequent extra-endosomal localization, as judged by live-cell imaging, proton, and electron microscopy, respectively. Even when released from heat-killed necrotic HEp-2 cells, GNP10 inhibited the necrotic tumor cell-induced maturation and functions of DCs, potentiated their Th2/Th17 polarization capacity, and thus, impaired the DCs' capacity to induce T cell-mediated anti-tumor cytotoxicity in vitro. Therefore, GNP10 could potentially induce more adverse DC-mediated immunological effects, compared to GNP50.
    Full-text · Article · May 2014 · PLoS ONE
  • [Show abstract] [Hide abstract]
    ABSTRACT: Royal jelly (RJ) fatty acids have recently been shown to possess various pharmacological and biological activities. In this work, we studied the immunomodulatory effects of 10-hydroxy-trans-2-decenoic acid (10-HDA) and 3,10-dihydroxy-decanoic acid (3,10-DDA), isolated from RJ, using a model of phytohaemagglutinin-activated human peripheral blood mononuclear cells (PBMCs). We showed that higher concentrations (500 μM) of both fatty acids inhibited the proliferation of PBMCs, and the process was followed by a decrease in the production of interleukin-2 (IL-2). 10-HDA at the concentration of 500 μM inhibited the production of IL-1β and tumor necrosis factor-α by stimulated PBMCs, whereas the same dose of 3,10-DDA had no effect on the levels of these cytokines. Regarding T helper (Th) cytokine profile, higher concentration of 10-HDA, in contrast to the lower one (50 μM), inhibited both Th1 and Th2 response, whereas Th17 response was not significantly modulated, as judged by the levels of interferon-γ, IL-5 and IL-17A in culture supernatants, respectively. Lower concentration of 3,10-DDA stimulated Th1 and Th17 responses and inhibited IL-10 production, whereas the higher dose augmented the Th2 response. In conclusion, our results showed a significant, dose-dependent, immunomodulatory effect of RJ fatty acids in vitro, which was also associated with their structure.
    No preview · Article · May 2014 · European Food Research and Technology
  • Source
    Vladimir Turuntas · Miodrag Colic · Sasa Vasilijic

    Full-text · Article · Feb 2014
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Immunoinflammatory mediated demyelination, the main pathological feature of multiple sclerosis (MS), is regularly accompanied by neurodegenerative processes, mostly in the form of axonal degeneration, which could be initiated by glutamate excitotoxicity. In the current study, the relationship between Th17 mediated inflammatory and excitotoxic events was investigated during an active phase of MS. Cerebrospinal fluid (CSF) of MS patients and control subjects was collected and IL-17A and glutamate levels were determined. IL-17A level was significantly higher in MS patients; whereas no statistically significant changes in glutamate concentrations were found. There was a direct correlation between IL-17A and glutamate levels; IL-17A levels were also associated with the neutrophil expansion in CSF and blood brain barrier disruption. However, IL-17A level and the number of neutrophils tended to fall with disease duration. The results suggest that Th17 cells might enhance and use glutamate excitotoxicity as an effector mechanism in the MS pathogenesis. Furthermore, Th17 immune response, as well as neutrophils, could be more important for MS onset rather than further disease development and progression, what could explain why some MS clinical trials, targeting Th17 cells in the later stage of the disease, failed to provide any clinical benefit. This article is protected by copyright. All rights reserved.
    Preview · Article · Jan 2014 · Scandinavian Journal of Immunology
  • Source

    Full-text · Article · Nov 2013 · European Journal of Vascular and Endovascular Surgery
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Nickel-chromium (Ni-Cr) dental alloys have been widely used in prosthodontic practice, but there is a permanent concern about their biocompatibility due to the release of metal ions. This is especially important when Ni-Cr metal microparticles are incorporated into gingival tissue during prosthodontic procedures. Therefore, the aim of this study was to examine and compare the corrosion and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy. Ni-Cr alloy, Remanium CSe bars (4 mm diameter), were made by the standard casting method and then cut into 0.5-mm-thick disks. Metal particles were obtained by scraping the bars using a diamond instrument for crown preparation. The microstructure was observed by an optical microscope. Quantitative determination and morphological and dimensional characterization of metal particles were carried out by a scanning electron microscope and Leica Application Suite software for image analysis. Corrosion was studied by conditioning the alloy specimens in the RPMI 1640 medium, containing 10% fetal calf serum in an incubator with 5% CO2 for 72 hours at 37°C. Inductively coupled plasma-optical emission spectrometry was used to assess metal ion release. The cytotoxity of conditioning medium (CM) was investigated on L929 cells using an MTT test. One-way ANOVA was used for statistical analysis. After casting, the microstructure of the Remanium CSe compact specimen composed of Ni, Cr, Mo, Si, Fe, Al, and Co had a typical dendritic structure. Alloy microparticles had an irregular shape with a wide size range: from less than 1 μm to more than 100 μm. The release of metal ions, especially Ni and Mo from microparticles, was significantly higher, compared to the compact alloy specimen. The CM prepared from compact alloy was not cytotoxic at any tested dilutions, whereas CM from alloy microparticles showed dose-dependent cytotoxicity (90% CM and 45% CM versus control; p < 0.005). Ni-Cr microparticles showed less corrosion resistance and lower biocompatibility than compact alloy. This could affect health on long-term exposure, especially in sensitized individuals.
    Full-text · Article · Oct 2013 · Journal of Prosthodontics
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: OBJECTIVES: ACST-2 is currently the largest trial ever conducted to compare carotid artery stenting (CAS) with carotid endarterectomy (CEA) in patients with severe asymptomatic carotid stenosis requiring revascularization. METHODS: Patients are entered into ACST-2 when revascularization is felt to be clearly indicated, when CEA and CAS are both possible, but where there is substantial uncertainty as to which is most appropriate. Trial surgeons and interventionalists are expected to use their usual techniques and CE-approved devices. We report baseline characteristics and blinded combined interim results for 30-day mortality and major morbidity for 986 patients in the ongoing trial up to September 2012. RESULTS: A total of 986 patients (687 men, 299 women), mean age 68.7 years (SD ± 8.1) were randomized equally to CEA or CAS. Most (96%) had ipsilateral stenosis of 70-99% (median 80%) with contralateral stenoses of 50-99% in 30% and contralateral occlusion in 8%. Patients were on appropriate medical treatment. For 691 patients undergoing intervention with at least 1-month follow-up and Rankin scoring at 6 months for any stroke, the overall serious cardiovascular event rate of periprocedural (within 30 days) disabling stroke, fatal myocardial infarction, and death at 30 days was 1.0%. CONCLUSIONS: Early ACST-2 results suggest contemporary carotid intervention for asymptomatic stenosis has a low risk of serious morbidity and mortality, on par with other recent trials. The trial continues to recruit, to monitor periprocedural events and all types of stroke, aiming to randomize up to 5,000 patients to determine any differential outcomes between interventions. Clinical trial: ISRCTN21144362.
    Full-text · Article · Sep 2013 · European Journal of Vascular and Endovascular Surgery

  • No preview · Conference Paper · Aug 2013
  • [Show abstract] [Hide abstract]
    ABSTRACT: The granulomatous foreign-body reaction (GFBR) is a special type of chronic inflammation characterised by infiltration of inflammatory cells (ICs). Using a model of subcutaneous implantation of polyvinyl sponges in rats, we studied the phenotypic and functional characteristics of these cells, with particular reference to the properties of antigen presenting cells and lymphocytes. ICs were isolated from sponge exudates at days 1-21 after implantation. Dendritic cells (DCs) and lymphocytes were purified using a combination of separation gradients, adherence to plastics, and immunomagnetic sorting. We showed that dominant population of ICs at day 1 and 3 were granulocytes, followed by their decrease thereafter. The number of mononuclear cells (MNCs), macrophages and DCs, progressively increased after day 3, reaching maximal values at day 7. Maximal number of lymphocytes was detected at day 10. In addition, total ICs isolated from day 7 till day 10 exerted significant suppressive activity in co-culture with autologous ConA-stimulated thymocytes and alogeneic lymph node T cells. This finding correlated with the increased level of IL-10 in culture supernatants and the increased proportion MHC class II+ IDO+ DCs and CD3+ CD25+ Foxp3+ lymphocytes at day 7 and day 10, respectively. We did not find any significant difference in the number of these cell populations between regional lymph nodes MNCs and peripheral blood MNCs of experimental animals compared to controls. In conclusion, accumulation of cells with tolerogenic characteristics in GFBR might be relevant for the suppression of inflammation and the prevention of unwanted immune response.
    No preview · Conference Paper · Aug 2013
  • [Show abstract] [Hide abstract]
    ABSTRACT: Introduction: Granulocytes are essential mediators of the acute inflammatory response and resolution of infection. Numerous studies showed that the function of these cells in children with cystic fibrosis (CF) is affected severely, although the mechanisms are not elucidated fully. Since Toll-like receptors (TLRs) mediate the microbicidal activity and inflammatory pathology of granulocytes, we wondered whether the TLR signaling in granulocytes of CF children is different from that of healthy donors. Materials and Methods: Granulocytes isolated from peripheral blood by density gradient, were cultivated either with inactivated Aspergillus conidia (a common infectious agent in CF), TLR agonists (TLR2-zymosan, TLR4-LPS, TLR9-CpG) or N-acetyl-cystein (used in CF therapy) for 2h, following the evaluation of myeloperoxidase activity by modified Graham-Knoll procedure. The apoptosis of granulocytes was evaluated after 24h-cultures by analyzing the chromatin condensation, whereas their cytokine production (IL-8 and IL-6) was assessed by ELISA. Results: Granulocytes of CF patients stimulated with Aspergillus, LPS, or zymosan had significantly lower myeloperoxidase activity compared to controls, and N-acetyl-cystein additionally diminished their myeloperoxidase activity. The apoptosis of granulocytes of CF patients was more inhibited by TLR stimuli and additionally potentiated by Aspergillus, compared to controls. IL-6 and IL-8 levels produced by granulocytes of CF patients was higher than in control cultures, especially in children with exacerbations or advanced stage of the disease (FEV1 and FVC<30%). Conclusion: By using different stimuli and inhibitors it might be possible to modify functional activity of granulocytes, and thus provide a potential target for modulating the immune response to Aspergillus fumigates and other pathogens in CF.
    No preview · Conference Paper · Jul 2013
  • [Show abstract] [Hide abstract]
    ABSTRACT: In the present study we investigated the cytochrome C oxidase (CO) activity and nitric oxide synthase (NOS) isoenzyme expression after intrahippocampal AlCl3 application in selective vulnerable brain structures. A single dose of AlCl3 was applied in the CA1 sector of rats hippocampus. For biochemical analysis, the animals were killed 10 min and three days after the treatment and forebrain cortex, basal forebrain and hippocampus were removed. Activity of CO was decreased bilaterally in the AlCl3-treated groups in all examined brain structures. We also applied immunohistochemical techniques to identify changes induced by AlCl3 injection after survival periods of 10 min and three days. Both the nNOS and eNOS stains were detected in the hippocampus of controls and AlCl3-treated animals, but iNOS labelling was present in the hippocampus only three days after AlCl3 application. An increased iNOS expression three days post AlCl3 administration could be involved in the mechanism of CO activity depletion.
    No preview · Article · Jul 2013

Publication Stats

2k Citations
286.05 Total Impact Points

Institutions

  • 2006-2015
    • University of Belgrade
      • Department of Microbiology and Immunology
      Beograd, Central Serbia, Serbia
  • 2011-2014
    • University of Niš
      • Faculty of Medicine (MF)
      Nisch, Central Serbia, Serbia
  • 1988-2014
    • Military Medical Academy
      Beograd, Central Serbia, Serbia
  • 2000-2013
    • Klinički centar Srbije
      • • Clinical Center of Serbia
      • • Institute for Cardiovascular Diseases
      Beograd, Central Serbia, Serbia
  • 2001-2012
    • Vojna akademija Beograd
      Beograd, Central Serbia, Serbia
  • 2006-2011
    • Institute for Educational Research, Belgrade, Serbia
      Beograd, Central Serbia, Serbia
  • 2009
    • University of Maribor
      • Faculty of Mechanical Engineering
      Maribor, Maribor, Slovenia
  • 2007
    • Institute for Educational Research, Belgrade, Serbia
      Beograd, Central Serbia, Serbia
  • 1999
    • Beograd Centar Medical Center for Maxillofacial Surgery
      Beograd, Central Serbia, Serbia
  • 1994
    • Institute for Medical Research - Belgrade
      Beograd, Central Serbia, Serbia