ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.

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epithelial barrier functions in immunocompetent individuals up to inflammatory diseases and severe life-threatening infections in immunocompromised patients. C. glabrata is of special importance because of a recent increase in its frequency and its innately reduced susceptibility to antifungal agents, specifically to azoles. The immunomodulation of fungal infections depends on the specific recognition of cell-wall antigens, representing the pathogen associated molecular patterns by immunocompetent cells. These antigens could be able to induce proliferation and activation of immune cells. The outermost layer of the cell wall of Candida species consists of mannoproteins containing O-glycosylated oligosaccharide and N-glycosylated polysaccharide moieties. Both carbohydrate moieties have been shown to play an essential role in host-fungal interactions, including the triggering and modulation of the anti-Candida host immune responses, which appear to rely on the interplay between the innate and adaptive immunities. Until yet, the role of mannan on immune system modulation is not sufficiently described. The dendritic cells (DCs) model represents a promising target for immunotherapy interventions and vaccine development. DCs increase an innate or specific response to fungi by producing inflammatory mediators, they are uniquely appropriate for decoding the fungus-associated information and next translation into a qualitatively different T helper responses. We observed, that C. glabrata mannan is able to induce activation of DCs and to increase the expression of co-stimulatory molecules CD80 and CD86. C. glabrata mannan was also able to stimulate proliferation of mouse splenocytes and increase production of TNF-α and IL-4 in concentration dependent manner. Acknowledgements This contribution is the result of project implementation: Centre of Excellence for Glycomics, ITMS 26240120031, supported by the Research & Development Operational Programme funded by ERDF. This work was supported by the Grant Agency of Slovak Academy of Sciences VEGA 2/0026/13 and by FEMS Research Fellowship FRF 2010-2. Legend to figure Fig.1: Expression of CD80 and CD86 DCs stimulated for 24 h with 10 μg/ml E. coli lipopolysaccharide (LPS10), 100μg/ml mannan C. glabrata (M100), 300μg/ml mannan C. glabrata (M300) and 600μg/ml mannan C. glabrata (M600). Analytical Cytometry VII 111
P21. ASSESSMENT OF MITOCHONDRIAL FUNCTIONS Barbora Pavlů, Jan Černý Faculty of Science, Charles University in Prague, Czech Republic; barbora.pavlu@natur.cuni.cz Mitochondria have become targets for activating specific killing of cancer cells and as a consequence this creates a need for assessment of drugs targeting mitochondria. To test effects of these substances - so called ”mitocans”( Ralph et al., 2006) - on mitochondria we employed MitoTracker ® Red CMXRos ( Molecular Probes® Invitrogen detection technologies), that is chloromethyl-X-rosamine. This fluorescent probe is retained in active mitochondria and its accumulation is dependent upon membrane potential. Results obtained with this mitochondrion selective probe by FACS analysis correspond with qualitative data from fluorescence microscopy. We found that working concentration for FACS analysis is between 2-5 nM. Suggested mechanism of induction of apoptosis by mitocans is a production of reactive oxygen species (ROS), which affect mitochondria in multiple ways (Neužil et al., 2006). To test production of ROS in cells treated by mitocans we used CellROX ® Green Reagent ( Molecular Probes® Invitrogen detection technologies). This probe is non-fluorescent in reduced state. After oxidation it becomes fluorescent and binds to DNA. Once again data from FACS analysis were in agreement with fluorescence microscopy. We found that working concentration for FACS analysis is between 10-30 µM. These two probes are compatible for use in multicolour experiments and are suitable for assessment of mitochondrial functions in cells treated by mitocans. Acknowledgements This work was supported by Grant Agency of the Czech Republic (P302/13/16565S), the Czech Ministry of Education, Youth and Sports of the Czech Republic (MSM0021620858) and by the grant SVV (265211). References Neuzil J, Wang XF, Dong LF, Low P, Ralph SJ.: Molecular mechanism of ‘mitocan’-induced apoptosis in cancer cells epitomizes the multiple roles of reactive oxygen species and Bcl-2 family proteins. - FEBS Lett. 2006 Oct 2;580(22):5125-9. Ralph SJ, Low P, Dong L, Lawen A, Neuzil J.: Mitocans: mitochondrial targeted anti-cancer drugs as improved therapies and related patent documents. - Recent Pat Anticancer Drug Discov. 2006 Nov;1(3):327-46. 112 Analytical Cytometry VII
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ABSTRACTS - ORAL PRESENTATIONS 14.
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25. PROFILING OF CHILDHOOD ACUTE LE
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29. INFLUENCE OF THE LEVEL OF CD133
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36. B-CELL IMMUNOPHENOTYPING OF LAR
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possible mechanism behind risk alle
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the frequency of autoimmune disease
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P65. OVULATION STIMULATION PROTOCOL
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Acknowledgements This work was supp
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P68. SUBPOPULATIONS OF B LYMPHOCYTE
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triggers a process of normal blood
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3 Department of Internal Medicine -
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P73. THE V-ATPASE-INHIBITOR BAFILOM
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P75. 5-FLUOROURACIL-SELECTED TUMOUR
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We conclude that decrease in the po
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Acknowledgements This work was supp
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P79. NEW TYPE OF PHOTOSENSITIZER IS
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LD11015, from GAČR 13-29358S, and
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given cell type are missing and the
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Our results show that both p38α +/
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for chemokines in attracting “inf
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ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.